Transgenic plants with enhanced agronomic traits

ABSTRACT

This invention provides transgenic plant cells with recombinant DNA for expression of proteins that are useful for imparting enhanced agronomic trait(s) to transgenic crop plants. This invention also provides transgenic plants and progeny seed comprising the transgenic plant cells where the plants are selected for having an enhanced trait selected from the group of traits consisting of enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil. Also disclosed are methods for manufacturing transgenic seed and plants with enhanced trait.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit under 35 USC §119(e) of U.S. provisional application Ser. Nos. 61/101,722, filed Oct. 1, 2008, 61/139,164, filed Dec. 19, 2008, 61/148,438, filed Jan. 30, 2009, 61/155,950, filed Feb. 23, 2009, 61/164,664, filed Mar. 30, 2009, 61/182,785, filed Jun. 1, 2009 and 61/226,953, filed Jul. 20, 2009, each of which is incorporated herein by reference in its entirety.

INCORPORATION OF SEQUENCE LISTING

The sequence listing file named “56202_B_seq_listing.txt”, which is 138,581,829 bytes (measured in MS-WINDOWS) which was electronically filed on a CD-ROM which was created on Sep. 28, 2009 is incorporated herein by reference.

FIELD OF THE INVENTION

Disclosed herein are recombinant DNA useful for providing enhanced traits to transgenic plants, seeds, pollen, plant cells and plant nuclei of such transgenic plants, methods of making and using such recombinant DNA, plants, seeds, pollen, plant cells and plant nuclei. Also disclosed are methods of producing hybrid corn seed comprising such recombinant DNA.

All genetic resources disclosed herein were directly obtained from sources that are currently common to the United States; the ancestral sources of each specific genetic material is unknown.

SUMMARY OF THE INVENTION

An aspect of this invention provides recombinant DNA constructs comprising polynucleotides characterized by an encoded protein having amino acids representing a protein family domain module as described in Table 10. Another aspect of this invention provides recombinant DNA constructs comprising polynucleotides characterized by an encoded protein with an amino acid sequence that is at least 90% identical to a corresponding consensus sequence defined in table 8. Yet another aspect of this invention provides recombinant DNA constructs comprising polynucleotides characterized by reference to SEQ ID NO:1-307 and the cognate proteins with amino acid sequences having reference to SEQ ID NO:308-614. The recombinant DNA constructs are useful for providing enhanced traits when stably integrated into the chromosomes and expressed in the nuclei of transgenic plants cells. In some aspects of the invention the recombinant DNA constructs, when expressed in a plant cell, provide for expression of cognate proteins. In those aspects of the invention, the recombinant DNA constructs for expressing cognate proteins are characterized by cognate amino acid sequences having a sequence selected from SEQ ID NOs: 308, 310, 312-315, 317-323, 325-343, 345, 347-349, 352-354, 356, 358-359, 366-372, 374-383, 389-392, 394, 396, 401-403, 405-412, 414, 417-424, 427-453, 455-473, 475, 488-501, 503-517, 519-531, 533-540, 542-543, and 546-614; having at least 90% identity over at least 90% of the length of a sequence selected from the group consisting of SEQ ID NOs: 308, 310, 312-315, 317-323, 325-343, 345, 347-349, 352-354, 356, 358-359, 366-372, 374-383, 389-392, 394, 396, 401-403, 405-412, 414, 417-424, 427-453, 455-473, 475, 488-501, 503-517, 519-531, 533-540, 542-543, and 546-614 or that are homologous to a sequence selected from the group consisting of SEQ ID NOs: 308, 310, 312-315, 317-323, 325-343, 345, 347-349, 352-354, 356, 358-359, 366-372, 374-383, 389-392, 394, 396, 401-403, 405-412, 414, 417-424, 427-453, 455-473, 475, 488-501, 503-517, 519-531, 533-540, 542-543, and 546-614.

In other aspects of the invention the recombinant DNA constructs provide for suppression of a native protein. In those other aspects of the invention the recombinant DNA constructs are characterized as being constructed with sense-oriented and anti-sense-oriented polynucleotides, e.g. polynucleotides derived from genes having SEQ ID NOs: 2, 4, 9, 17, 37, 39, 43-44, 48, 50, 53-58, 66, 77-81, 86, 88, 90-93, 97, 106, 108-109, 118-119, 147, 167, 169-180, 195, 211, 225, 234, or 237-238 or homologous genes. When the recombinant DNA construct is expressed in corn plants, the endogenous protein is a corn protein with an amino acid sequence of SEQ ID NO:316, 344, 346, 350-351, 355, 357, 360-365, 384-388, 393, 397-400, 404, 413, 415-416, 425-426, 474, 476-487, 502, 532, 541, or 544-545 or the corn homolog of SEQ ID NOs:309, 311, 324, 373, 395, 454, or 518; when the recombinant DNA construct is expressed in soybean plants, the endogenous protein is a soybean protein with an amino acid sequence of SEQ ID NO: 309, 324, 373, 395, 518 or is a soybean homolog of SEQ ID NOs: 311, 316, 344, 346, 350-351, 355, 357, 360-365, 384-388, 393, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 532, 541, or 544-545; and when the recombinant DNA construct is expressed in a plant other than a corn or a soybean plant, the endogenous protein is the other plant's endogenous protein that has an amino acid sequence homologous to SEQ ID NO: 309, 311, 316, 324, 344, 346, 350-351, 355, 357, 360-365, 373, 384-388, 393, 395, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 518, 532, 541, or 544-545.

In practical aspects of this invention the recombinant DNA constructs of the invention are stably integrated into the chromosome of a plant cell nucleus.

This invention also provides transgenic plant cells comprising the stably integrated recombinant DNA constructs of the invention, transgenic plants and seeds comprising a plurality of such transgenic plant cells and transgenic pollen of such plants. Such transgenic plants are selected from a population of transgenic plants regenerated from plant cells transformed with recombinant DNA constructs by screening transgenic plants for an enhanced trait as compared to control plants. The enhanced trait is one or more of enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil.

In another aspect of the invention the plant cells, plants, seeds, and pollen further comprise DNA expressing a protein that provides tolerance from exposure to an herbicide applied at levels that are lethal to a wild type plant cell.

This invention also provides methods for manufacturing non-natural, transgenic seed that can be used to produce a crop of transgenic plants with an enhanced trait resulting from expression of a stably-integrated recombinant DNA construct. More specifically, the method comprises (a) screening a population of plants for an enhanced trait and a recombinant DNA construct, where individual plants in the population can exhibit the trait at a level less than, essentially the same as or greater than the level that the trait is exhibited in control plants, (b) selecting from the population one or more plants that exhibit the trait at a level greater than the level that said trait is exhibited in control plants, (c) collecting seed from a selected plant, (d) verifying that the recombinant DNA is stably integrated in said selected plants, (e) analyzing tissue of a selected plant to determine the production or suppression of a protein having the function of a protein encoded by nucleotides in a sequence of one of SEQ ID NOs:1-307. In one aspect of the invention, the plants in the population further comprise DNA expressing a protein that provides tolerance to exposure to a herbicide applied at levels that are lethal to wild type plant cells and the selecting is affected by treating the population with the herbicide, e.g. a glyphosate, dicamba, or glufosinate compound. In another aspect of the invention the plants are selected by identifying plants with the enhanced trait. The methods are especially useful for manufacturing corn, soybean, cotton, canola, alfalfa, wheat, rice, sugarcane or sugar beet seed.

Another aspect of the invention provides a method of producing hybrid corn seed comprising acquiring hybrid corn seed from a herbicide tolerant corn plant which also has stably-integrated, recombinant DNA construct comprising a promoter that is (a) functional in plant cells and (b) is operably linked to DNA that encodes or suppresses a protein having the function of a protein encoded by nucleotides in a sequence of one of SEQ ID NOs:1-307. The methods further comprise producing corn plants from said hybrid corn seed, wherein a fraction of the plants produced from said hybrid corn seed is homozygous for said recombinant DNA, a fraction of the plants produced from said hybrid corn seed is hemizygous for said recombinant DNA, and a fraction of the plants produced from said hybrid corn seed has none of said recombinant DNA; selecting corn plants which are homozygous and hemizygous for said recombinant DNA by treating with an herbicide; collecting seed from herbicide-treated-surviving corn plants and planting said seed to produce further progeny corn plants; repeating the selecting and collecting steps at least once to produce an inbred corn line; and crossing the inbred corn line with a second corn line to produce hybrid seed.

Another aspect of the invention provides a method of selecting a plant comprising plant cells of the invention by using an immunoreactive antibody to detect the presence or absence of protein expressed or suppressed by recombinant DNA in seed or plant tissue. Yet another aspect of the invention provides anti-counterfeit milled seed having, as an indication of origin, plant cells of this invention.

Still other aspects of this invention relate to transgenic plants with enhanced water use efficiency or enhanced nitrogen use efficiency. For instance, this invention provides methods of growing a corn, cotton, soybean, or canola crop without irrigation water comprising planting seed having plant cells of the invention which are selected for enhanced water use efficiency. Alternatively methods comprise applying reduced irrigation water, e.g. providing up to 300 millimeters of ground water during the production of a corn crop. This invention also provides methods of growing a corn, cotton, soybean or canola crop without added nitrogen fertilizer comprising planting seed having plant cells of the invention which are selected for enhanced nitrogen use efficiency.

DETAILED DESCRIPTION OF THE INVENTION

In the attached sequence listing:

SEQ ID NO:1-307 are nucleotide sequences of the coding strand of DNA for “genes” used in the recombinant DNA imparting an enhanced trait in plant cells, i.e. each represents a coding sequence for a protein;

SEQ ID NO: 308-614 are amino acid sequences of the cognate protein of the “genes” with nucleotide coding sequences 1-307;

SEQ ID NO: 615-36442 are amino acid sequences of homologous proteins;

SEQ ID NO: 36443 is a nucleotide sequence of a base plasmid vector useful for corn transformation;

SEQ ID NO: 36444 is a nucleotide sequence of a base plasmid vector useful for soybean and canola transformation;

SEQ ID NO: 36445 is a nucleotide sequence of a base plasmid vector useful for cotton transformation;

SEQ ID NO: 36446 is a nucleotide sequence of a base plasmid vector useful for co-transformation to produce gene stacks in corn;

SEQ ID NO: 36447-36478 are consensus amino acid sequences. Table 8 lists the protein SEQ ID NOs and their corresponding consensus SEQ ID NOs.

As used herein a “plant cell” means a plant cell that is transformed with stably-integrated, non-natural, recombinant DNA, e.g. by Agrobacterium-mediated transformation or by bombardment using microparticles coated with recombinant DNA or other means. A plant cell of this invention can be an originally-transformed plant cell that exists as a microorganism or as a progeny plant cell that is regenerated into differentiated tissue, e.g. into a transgenic plant with stably-integrated, non-natural recombinant DNA, or seed or pollen derived from a progeny transgenic plant.

As used herein a “transgenic plant” means a plant whose genome has been altered by the stable integration of recombinant DNA. A transgenic plant includes a plant regenerated from an originally-transformed plant cell and progeny transgenic plants from later generations or crosses of a transformed plant.

As used herein “recombinant DNA” means DNA which has been a genetically engineered and constructed outside of a cell including DNA containing naturally occurring DNA or cDNA or synthetic DNA.

As used herein “consensus sequence” means an artificial sequence of amino acids in a conserved region of an alignment of amino acid sequences of homologous proteins, e.g. as determined by a CLUSTALW alignment of amino acid sequence of homolog proteins.

As used herein a “homolog” means a protein in a group of proteins that perform the same biological function, e.g. proteins that belong to the same Pfam protein family and that provide a common enhanced trait in transgenic plants of this invention. Homologs are expressed by homologous genes. With reference to homologous genes, homologs include orthologs, i.e. genes expressed in different species that evolved from a common ancestral genes by speciation and encode proteins retain the same function, but do not include paralogs, i.e. genes that are related by duplication but have evolved to encode proteins with different functions. Homologous genes include naturally occurring alleles and artificially-created variants. Degeneracy of the genetic code provides the possibility to substitute at least one base of the protein encoding sequence of a gene with a different base without causing the amino acid sequence of the polypeptide produced from the gene to be changed. When optimally aligned, homolog proteins have at least 60% identity, 65% identity, 70% identity, 75% identity, 80%, identity, 85% identity, 90% identity, 95, 96, 97, 98, or 99% identity over the full length of a protein identified as being associated with imparting an enhanced trait when expressed in plant cells. In one aspect of the invention homolog proteins have an amino acid sequence that has at least 90% identity to a consensus amino acid sequence of proteins and homologs disclosed herein.

Homologs are identified by comparison of amino acid sequence, e.g. manually or by use of a computer-based tool using known homology-based search algorithms such as the suite of BLAST programs available from NCBI. A local sequence alignment program, e.g. BLAST, can be used to search a database of sequences to find similar sequences, and the summary Expectation value (E-value) used to measure the sequence base similarity. Because a protein hit with the best E-value for a particular organism may not necessarily be an ortholog, i.e. have the same function, or be the only ortholog, a reciprocal query is used to filter hit sequences with significant E-values for ortholog identification. The reciprocal query entails search of the significant hits against a database of amino acid sequences from the base organism that are similar to the sequence of the query protein. A hit can be identified as an ortholog, when the reciprocal query's best hit is the query protein itself or a protein encoded by a duplicated gene after speciation. A further aspect of the homologs encoded by DNA useful in the transgenic plants of the invention are those proteins that differ from a disclosed protein as the result of deletion or insertion of one or more amino acids in a native sequence.

Percent identity describes the extent to which the sequences of DNA or protein segments are invariant in an alignment of sequences, for example nucleotide sequences or amino acid sequences. An alignment of sequences is created by manually aligning two sequences, e.g. a stated sequence, as provided herein, as a reference, and another sequence, to produce the highest number of matching elements, e.g. individual nucleotides or amino acids, while allowing for the introduction of gaps into either sequence. An “identity fraction” for a sequence aligned with a reference sequence is the number of matching elements, divided by the full length of the reference sequence, not including gaps introduced by the alignment process into the reference sequence. “Percent identity” (“% identity”) as used herein is the identity fraction times 100.

“Pfam” is a large collection of multiple sequence alignments and hidden Markov models covering many common protein families, e.g. Pfam version 19.0 (December 2005) contains alignments and models for 8183 protein families and is based on the Swissprot 47.0 and SP-TrEMBL 30.0 protein sequence databases. See S. R. Eddy, “Profile Hidden Markov Models”, Bioinformatics 14:755-763, 1998. The Pfam database is currently maintained and updated by the Pfam Consortium. The alignments represent some evolutionary conserved structure that has implications for the proteids function. Profile hidden Markov models (profile HMMs) built from the protein family alignments are useful for automatically recognizing that a new protein belongs to an existing protein family even if the homology by alignment appears to be low.

Protein domains are identified by querying the amino acid sequence of a protein against Hidden Markov Models which characterize protein family domains (“Pfam domains”) using HMMER software, which is available from the Pfam Consortium. The HMMER software is also disclosed in patent application publication US 2008/0148432 A1 incorporated herein by reference. A protein domain meeting the gathering cutoff for the alignment of a particular Pfam domain is considered to contain the Pfam domain.

A “Pfam domain module” is a representation of Pfam domains in a protein, in order from N terminus to C terminus. In a Pfam domain module individual Pfam domains are separated by double colons “::”. The order and copy number of the Pfam domains from N to C terminus are attributes of a Pfam domain module. Although the copy number of repetitive domains is important, varying copy number often enables a similar function. Thus, a Pfam domain module with multiple copies of a domain should define an equivalent Pfam domain module with variance in the number of multiple copies. A Pfam domain module is not specific for distance between adjacent domains, but contemplates natural distances and variations in distance that provide equivalent function. The Pfam database contains both narrowly- and broadly-defined domains, leading to identification of overlapping domains on some proteins. A Pfam domain module is characterized by non-overlapping domains. Where there is overlap, the domain having a function that is more closely associated with the function of the protein (based on the E value of the Pfam match) is selected.

Once one DNA is identified as encoding a protein which imparts an enhanced trait when expressed in transgenic plants, other DNA encoding proteins with the same Pfam domain module are identified by querying the amino acid sequence of protein encoded by candidate DNA against the Hidden Markov Models which characterizes the Pfam domains using HMMER software. Candidate proteins meeting the same Pfam domain module are in the protein family and have cognate DNA that is useful in constructing recombinant DNA for the use in the plant cells of this invention. Hidden Markov Model databases for use with HMMER software in identifying DNA expressing protein with a common Pfam domain module for recombinant DNA in the plant cells of this invention are available from the Pfam Consortium (ftp.sanger.ac.uk/pub/databases/Pfam/) and are incorporated herein by reference.

The HMMER software and Pfam databases (version 23.0) were used to identify known domains in the proteins corresponding to amino acid sequence of SEQ ID NOs: 308-310, 312-313, 315, 317-332, 334-343, 345, 347-364, 366-372, 374, 382, 387-403, 406-412, 414-425, 427-448, 450-460, 462-465, 467-477, 479-482, 484-487, 493-511, 513-539, 542, 545-558, 560-578, 580-582, 584-598, 600-602, 604-608, 612-614. All DNA encoding proteins that have scores higher than the gathering cutoff disclosed in Table 11 by Pfam analysis disclosed herein can be used in recombinant DNA of the plant cells of this invention, e.g. for selecting transgenic plants having enhanced agronomic traits. The relevant Pfams modules for use in this invention, as more specifically disclosed below, are PHD::SET, CBFB_NFYA, 2-Hacid_dh_C, 60KD_IMP, A_thal_(—)3526, AA_kinase, AA_kinase::NAD_binding_(—)3::Homoserine_dh, AA_permease, Aa_trans, ABC_membrane::ABC_tran, ABC_tran, Acetate_kinase, Acid_phosphat_B, Acyl-ACP_TE, AlaDh_PNT_N::AlaDh_PNT_C, Amino_oxidase, Aminotran_(—)1_(—)2 Aminotran_(—)5, Ammonium_transp, Ank::Ank::Ank::Ank::Ank::Ank::Ank::Ank, AP2, Arginase, Arginosuc_synth, AsnA, Asp, Asp_decarbox, ATP-sulfurylase, Auxin_inducible, BCCT, BPD_transp_(—)2, BTB, bZIP_(—)1, bZIP_(—)1::MethyltransfD12, bZIP_(—)2, bZIP_(—)2::bZIP_(—)1, C4dic_mal_tran, CBFB_NFYA, CCT, Clp_N::Clp_N::AAA::AAA_(—)2::ClpB_D2-small, cNMP_binding::Crp, CSD, DEAD::Helicase_C, DEAD_(—)2::DUF1227, Dehydrin Dimerisation::Methyltransf_(—)2, DSPc, DUF1292, DUF506, DUF640, DUF647, DUF828::PH_(—)2, eIF-5a, eRF1_(—)1::eRF1_(—)2::eRF1_(—)3, FAD_binding_(—)3::FHA, FAD_binding_(—)4::ALO, FAD_binding_(—)4::Lact-deh-memb, F-box::Tub, Fe-ADH, Form_Nir_trans, FTCD_N, Gal_Lectin, GATase_(—)2::Asn_synthase, GATase_(—)2::Glu_syn_centrat:Glu_synthase::GXGXG, GDPD, GH3, Gln-synt_N::Gln-synt_C, Globin::FAD_binding_(—)6::NAD_binding_(—)1, Glt_symporter, Glutaredoxin, Glutaredoxin::Glutaredoxin::Glutaredoxin, Glyco_transf_(—)20::Trehalose_PPase, GTP1_OBG::MMR_HSR1::DUF1967, HLH, HMG_box Homeobox::HALZ, Homeobox:: START, HR_lesion, HSP20, HSP70, LIM, LIM::LIM, LisH::WD40::WD40::WD40::WD40::WD40::WD40::WD40 LRR_(—)1::LRR_(—)1::LRR_(—)1::LRR_(—)1::Pkinase_Tyr, LRRNT_(—)2::LRR_(—)1::LRR_(—)1::LRR_(—)1::LRR_(—)1::LRR_(—)1::LRR_(—)1::LRR_(—)1::LRR_(—)1::LRR_(—)1::L RR_(—)1::LRR_(—)1::LRR_(—)1::LRR_(—)1::Pkinase, LRRNT_(—)2::LRR_(—)1::Pkinase_Tyr, LysM, MBD, MethyltransfD 12. MIT::AAA::Vps4_C, Mito_carr::Mito_carr::Mito_carr, MMR_HSR1::KH_(—)2, Monooxygenase_B, Myb_DNA-binding::Myb_DNA-binding, NAD_binding_(—)1, NAD_binding_(—)2, NAM, Ndr, NIR_SIR_ferr::NIR_SIR::NIR_SIR_ferr, Nitroreductase, NUDIX, OKR_DC_(—)1::OKR_DC_(—)1_C, OPT, p450, PAS_(—)2::GAF::Phytochrome::PAS::PAS::HisKA::HATPase_c, PBP, Peptidase_C2::Calpain_III, Peptidase_(—)510, peroxidase, Pkinase, Pkinase::NAF, PLAC8, PMEI, PDX::Homeobox, PP2C, PTA_PTB, PTR2 RCC1::RCC1::RCC1, Response_reg::Myb_DNA-binding, Ribosomal_L21p, RolB_RolC::Amino_oxidase, RRM_(—)1, RRM_(—)1::RRM_(—)1, RRM_(—)1::zf-CCHC RWP-RK::PB1, SBP, SBP_bac_(—)3, Sina, SIS::CBS, SOUL, SPRY, SRF-TF, SRF-TF::K-box, SSF, Ssl1::C1_(—)4, Sulfate_transp::STAS, Thg1::Thg1 Thiolase_N::Thiolase_C, Thioredoxin::Glutaredoxin::Glutaredoxin::Glutaredoxin, TPP_enzyme_N::TPP_enzyme_M::TPP_enzyme_C Transaldolase, tRNA_synt_(—)1c_R1::tRNA_synt_(—)1c_R2::tRNA-synt_(—)1c::tRNA-synt_(—)1c_C, ubiquitin, U-box, Usp, WD40::WD40, WRKY, WRKY::WRKY zf-B_box, zf-B_box::MethyltransfD12, zf-B_box::zf-B_box::CCT, zf-C3HC4, zf-CCCH, zf-CCHC::Plus-3, and zf-D of for which databases are included in the appended computer listing.

As used herein “promoter” means regulatory DNA for initializing transcription. A “plant promoter” is a promoter capable of initiating transcription in plant cells whether or not its origin is a plant cell, e.g. is it well known that Agrobacterium promoters are functional in plant cells. Thus, plant promoters include promoter DNA obtained from plants, plant viruses and bacteria such as Agrobacterium and Bradyrhizobium bacteria. Examples of promoters under developmental control include promoters that preferentially initiate transcription in certain tissues, such as leaves, roots, or seeds. Such promoters are referred to as “tissue preferred”. Promoters that initiate transcription only in certain tissues are referred to as “tissue specific”. A “cell type” specific promoter primarily drives expression in certain cell types in one or more organs, for example, vascular cells in roots or leaves. An “inducible” or “repressible” promoter is a promoter which is under environmental control. Examples of environmental conditions that may effect transcription by inducible promoters include anaerobic conditions, or certain chemicals, or the presence of light. Tissue specific, tissue preferred, cell type specific, and inducible promoters constitute the class of “non-constitutive” promoters. A “constitutive” promoter is a promoter which is active under most conditions.

As used herein “operably linked” means the association of two or more DNA fragments in a recombinant DNA construct so that the function of one, e.g. protein-encoding DNA, is controlled by the other, e.g. a promoter.

As used herein “expressed” means produced, e.g. a protein is expressed in a plant cell when its cognate DNA is transcribed to mRNA that is translated to the protein.

As used herein “suppressed” means decreased, e.g. a protein is suppressed in a plant cell when there is a decrease in the amount and/or activity of the protein in the plant cell. The presence or activity of the protein can be decreased by any amount up to and including a total loss of protein expression and/or activity.

As used herein a “control plant” means a plant that does not contain the recombinant DNA that imparts an enhanced trait. A control plant is used to identify and select a transgenic plant that has an enhanced trait. A suitable control plant can be a non-transgenic plant of the parental line used to generate a transgenic plant, i.e. devoid of recombinant DNA. A suitable control plant may in some cases be a progeny of a hemizygous transgenic plant line that does not contain the recombinant DNA, known as a negative segregant.

As used herein an “enhanced trait” means a characteristic of a transgenic plant that includes, but is not limited to, an enhance agronomic trait characterized by enhanced plant morphology, physiology, growth and development, yield, nutritional enhancement, disease or pest resistance, or environmental or chemical tolerance. In more specific aspects of this invention enhanced trait is selected from group of enhanced traits consisting of enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil. In an important aspect of the invention the enhanced trait is enhanced yield including increased yield under non-stress conditions and increased yield under environmental stress conditions. Stress conditions may include, for example, drought, shade, fungal disease, viral disease, bacterial disease, insect infestation, nematode infestation, cold temperature exposure, heat exposure, osmotic stress, reduced nitrogen nutrient availability, reduced phosphorus nutrient availability and high plant density. “Yield” can be affected by many properties including without limitation, plant height, pod number, pod position on the plant, number of internodes, incidence of pod shatter, grain size, efficiency of nodulation and nitrogen fixation, efficiency of nutrient assimilation, resistance to biotic and abiotic stress, carbon assimilation, plant architecture, resistance to lodging, percent seed germination, seedling vigor, and juvenile traits. Yield can also be affected by efficiency of germination (including germination in stressed conditions), growth rate (including growth rate in stressed conditions), ear number, seed number per ear, seed size, composition of seed (starch, oil, protein) and characteristics of seed fill.

Increased yield of a transgenic plant of the present invention can be measured in a number of ways, including test weight, seed number per plant, seed weight, seed number per unit area (i.e. seeds, or weight of seeds, per acre), bushels per acre, tons per acre, or kilo per hectare. For example, corn yield may be measured as production of shelled corn kernels per unit of production area, for example in bushels per acre or metric tons per hectare, often reported on a moisture adjusted basis, for example at 15.5 percent moisture. Increased yield may result from improved utilization of key biochemical compounds, such as nitrogen, phosphorous and carbohydrate, or from improved responses to environmental stresses, such as cold, heat, drought, salt, and attack by pests or pathogens. Recombinant DNA used in this invention can also be used to provide plants having improved growth and development, and ultimately increased yield, as the result of modified expression of plant growth regulators or modification of cell cycle or photosynthesis pathways. Also of interest is the generation of transgenic plants that demonstrate enhanced yield with respect to a seed component that may or may not correspond to an increase in overall plant yield. Such properties include enhancements in seed oil, seed molecules such as protein and starch, oil components as may be manifest by an alterations in the ratios of seed components.

Recombinant DNA constructs are assembled using methods well known to persons of ordinary skill in the art and typically comprise a promoter operably linked to DNA, the expression of which provides the enhanced agronomic trait. Other construct components may include additional regulatory elements, such as 5′ leaders and introns for enhancing transcription, 3′ untranslated regions (such as polyadenylation signals and sites), DNA for transit or signal peptides.

Numerous promoters that are active in plant cells have been described in the literature. These include promoters present in plant genomes as well as promoters from other sources, including nopaline synthase (NOS) promoter and octopine synthase (OCS) promoters carried on tumor-inducing plasmids of Agrobacterium tumefaciens and the CaMV35S promoters from the cauliflower mosaic virus as disclosed in U.S. Pat. Nos. 5,164,316 and 5,322,938. Useful promoters derived from plant genes are found in U.S. Pat. No. 5,641,876 which discloses a rice actin promoter, U.S. Pat. No. 7,151,204 which discloses a maize chloroplast aldolase promoter and a maize aldolase (FDA) promoter, and US Patent Application Publication 2003/0131377 A1 which discloses a maize nicotianamine synthase promoter. These and numerous other promoters that function in plant cells are known to those skilled in the art and available for use in recombinant polynucleotides of the present invention to provide for expression of desired genes in transgenic plant cells.

Furthermore, the promoters may be altered to contain multiple “enhancer sequences” to assist in elevating gene expression. Such enhancers are known in the art. By including an enhancer sequence with such constructs, the expression of the selected protein may be enhanced. These enhancers often are found 5′ to the start of transcription in a promoter that functions in eukaryotic cells, but can often be inserted upstream (5′) or downstream (3′) to the coding sequence. In some instances, these 5′ enhancing elements are introns. Particularly useful as enhancers are the 5′ introns of the rice actin 1 (see U.S. Pat. No. 5,641,876) and rice actin 2 genes, the maize alcohol dehydrogenase gene intron, the maize heat shock protein 70 gene intron (U.S. Pat. No. 5,593,874) and the maize shrunken 1 gene. See also US Patent Application Publication 2002/0192813A1 which discloses 5′, 3′ and intron elements useful in the design of effective plant expression vectors.

In other aspects of the invention, sufficient expression in plant seed tissues is desired to affect improvements in seed composition. Exemplary promoters for use for seed composition modification include promoters from seed genes such as napin as disclosed in U.S. Pat. No. 5,420,034, maize L3 oleosin as disclosed in U.S. Pat. No. 6,433,252), zein Z27 as disclosed by Russell et al. (1997) Transgenic Res. 6(2):157-166), globulin 1 as disclosed by Belanger et al (1991) Genetics 129:863-872), glutelin 1 as disclosed by Russell (1997) supra), and peroxiredoxin antioxidant (Per1) as disclosed by Stacy et al. (1996) Plant Mol. Biol. 31(6):1205-1216.

Recombinant DNA constructs useful in this invention will also generally include a 3′ element that typically contains a polyadenylation signal and site. Well-known 3′ elements include those from Agrobacterium tumefaciens genes such as nos 3′, tml 3′, tmr 3′, tms 3′, ocs 3′, tr7 3′, for example disclosed in U.S. Pat. No. 6,090,627; 3′ elements from plant genes such as wheat (Triticum aesevitum) heat shock protein 17 (Hsp17 3′), a wheat ubiquitin gene, a wheat fructose-1,6-biphosphatase gene, a rice glutelin gene, a rice lactate dehydrogenase gene and a rice beta-tubulin gene, all of which are disclosed in US Patent Application Publication 2002/0192813 A1; and the pea (Pisum sativum) ribulose biphosphate carboxylase gene (rbs 3′), and 3′ elements from the genes within the host plant.

Constructs and vectors may also include a transit peptide for targeting of a gene to a plant organelle, particularly to a chloroplast, leucoplast or other plastid organelle. For descriptions of the use of chloroplast transit peptides see U.S. Pat. No. 5,188,642 and U.S. Pat. No. 5,728,925. For description of the transit peptide region of an Arabidopsis EPSPS gene useful in the present invention, see Klee, H. J. et al (MGG (1987) 210:437-442).

Recombinant DNA constructs for gene suppression can be designed for any of a number the well-known methods for suppressing transcription of a gene, the accumulation of the mRNA corresponding to that gene or preventing translation of the transcript into protein. Posttranscriptional gene suppression can be practically effected by transcription of RNA that forms double-stranded RNA (dsRNA) having homology to mRNA produced from a gene targeted for suppression.

Gene suppression can also be achieved by insertion mutations created by transposable elements may also prevent gene function. For example, in many dicot plants, transformation with the T-DNA of Agrobacterium may be readily achieved and large numbers of transformants can be rapidly obtained. Also, some species have lines with active transposable elements that can efficiently be used for the generation of large numbers of insertion mutations, while some other species lack such options. Mutant plants produced by Agrobacterium or transposon mutagenesis and having altered expression of a polypeptide of interest can be identified using the polynucleotides of the present invention. For example, a large population of mutated plants may be screened with polynucleotides encoding the polypeptide of interest to detect mutated plants having an insertion in the gene encoding the polypeptide of interest.

Transgenic plants may comprise a stack of one or more polynucleotides disclosed herein resulting in the production or suppression of multiple polypeptide sequences. Transgenic plants comprising stacks of polynucleotide sequences can be obtained by either or both of traditional breeding methods or through genetic engineering methods. These methods include, but are not limited to, breeding individual lines each comprising a polynucleotide of interest, transforming a transgenic plant comprising a gene disclosed herein with a subsequent gene, and co-transformation of genes into a single plant cell. Co-transformation of genes can be carried out using single transformation vectors comprising multiple genes or genes carried separately on multiple vectors.

Transgenic plants comprising or derived from plant cells of this invention transformed with recombinant DNA can be further enhanced with stacked traits, e.g. a crop plant having an enhanced trait resulting from expression of DNA disclosed herein in combination with herbicide and/or pest resistance traits. For example, genes of the current invention can be stacked with other traits of agronomic interest, such as a trait providing herbicide resistance, or insect resistance, such as using a gene from Bacillus thuringensis to provide resistance against lepidopteran, coliopteran, homopteran, hemiopteran, and other insects. Herbicides for which transgenic plant tolerance has been demonstrated and the method of the present invention can be applied include, but are not limited to, glyphosate, dicamba, glufosinate, sulfonylurea, bromoxynil and norflurazon herbicides. Polynucleotide molecules encoding proteins involved in herbicide tolerance are well-known in the art and include, but are not limited to, a polynucleotide molecule encoding 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) disclosed in U.S. Pat. Nos. 5,094,945; 5,627,061; 5,633,435 and 6,040,497 for imparting glyphosate tolerance; polynucleotide molecules encoding a glyphosate oxidoreductase (GOX) disclosed in U.S. Pat. No. 5,463,175 and a glyphosate-N-acetyl transferase (GAT) disclosed in US Patent Application Publication 2003/0083480 A1 also for imparting glyphosate tolerance; dicamba monooxygenase disclosed in US Patent Application Publication 2003/0135879 A1 for imparting dicamba tolerance; a polynucleotide molecule encoding bromoxynil nitrilase (Bxn) disclosed in U.S. Pat. No. 4,810,648 for imparting bromoxynil tolerance; a polynucleotide molecule encoding phytoene desaturase (crtl) described in Misawa et al, (1993) Plant J. 4:833-840 and in Misawa et al, (1994) Plant J. 6:481-489 for norflurazon tolerance; a polynucleotide molecule encoding acetohydroxyacid synthase (AHAS, aka ALS) described in Sathasiivan et al. (1990) Nucl. Acids Res. 18:2188-2193 for imparting tolerance to sulfonylurea herbicides; polynucleotide molecules known as bar genes disclosed in DeBlock, et al. (1987) EMBO J. 6:2513-2519 for imparting glufosinate and bialaphos tolerance; polynucleotide molecules disclosed in US Patent Application Publication 2003/010609 A1 for imparting N-amino methyl phosphonic acid tolerance; polynucleotide molecules disclosed in U.S. Pat. No. 6,107,549 for impartinig pyridine herbicide resistance; molecules and methods for imparting tolerance to multiple herbicides such as glyphosate, atrazine, ALS inhibitors, isoxoflutole and glufosinate herbicides are disclosed in U.S. Pat. No. 6,376,754 and US Patent Application Publication 2002/0112260. Molecules and methods for imparting insect/nematode/virus resistance are disclosed in U.S. Pat. Nos. 5,250,515; 5,880,275; 6,506,599; 5,986,175 and US Patent Application Publication 2003/0150017 A1.

Plant Cell Transformation Methods

Numerous methods for transforming chromosomes in a plant cell nucleus with recombinant DNA are known in the art and are used in methods of preparing a transgenic plant cell nucleus cell, and plant. Two effective methods for such transformation are Agrobacterium-mediated transformation and microprojectile bombardment. Microprojectile bombardment methods are illustrated in U.S. Pat. Nos. 5,015,580 (soybean); 5,550,318 (corn); 5,538,880 (corn); 5,914,451 (soybean); 6,160,208 (corn); 6,399,861 (corn); 6,153,812 (wheat) and 6,365,807 (rice) and Agrobacterium-mediated transformation is described in U.S. Pat. Nos. 5,159,135 (cotton); 5,824,877 (soybean); 5,463,174 (canola); 5,591,616 (corn); 5,846,797 (cotton); 6,384,301 (soybean), 7,026,528 (wheat) and 6,329,571 (rice), US Patent Application Publication 2004/0087030 A1 (cotton), and US Patent Application Publication 2001/0042257 A1 (sugar beet), all of which are incorporated herein by reference for enabling the production of transgenic plants. Transformation of plant material is practiced in tissue culture on a nutrient media, i.e. a mixture of nutrients that will allow cells to grow in vitro. Recipient cell targets include, but are not limited to, meristem cells, hypocotyls, calli, immature embryos and gametic cells such as microspores, pollen, sperm and egg cells. Callus may be initiated from tissue sources including, but not limited to, immature embryos, hypocotyls, seedling apical meristems, microspores and the like. Cells containing a transgenic nucleus are grown into transgenic plants.

In addition to direct transformation of a plant material with a recombinant DNA, a transgenic plant cell nucleus can be prepared by crossing a first plant having cells with a transgenic nucleus with recombinant DNA with a second plant lacking the transgenic nucleus. For example, recombinant DNA can be introduced into a nucleus from a first plant line that is amenable to transformation to transgenic nucleus in cells that are grown into a transgenic plant which can be crossed with a second plant line to introgress the recombinant DNA into the second plant line. A transgenic plant with recombinant DNA providing an enhanced trait, e.g. enhanced yield, can be crossed with transgenic plant line having other recombinant DNA that confers another trait, for example herbicide resistance or pest resistance, to produce progeny plants having recombinant DNA that confers both traits. Typically, in such breeding for combining traits the transgenic plant donating the additional trait is a male line and the transgenic plant carrying the base traits is the female line. The progeny of this cross will segregate such that some of the plants will carry the DNA for both parental traits and some will carry DNA for one parental trait; such plants can be identified by markers associated with parental recombinant DNA, e.g. marker identification by analysis for recombinant DNA or, in the case where a selectable marker is linked to the recombinant, by application of the selecting agent such as a herbicide for use with a herbicide tolerance marker, or by selection for the enhanced trait. Progeny plants carrying DNA for both parental traits can be crossed back into the female parent line multiple times, for example usually 6 to 8 generations, to produce a progeny plant with substantially the same genotype as one original transgenic parental line but for the recombinant DNA of the other transgenic parental line

In the practice of transformation DNA is typically introduced into only a small percentage of target plant cells in any one transformation experiment. Marker genes are used to provide an efficient system for identification of those cells that are stably transformed by receiving and integrating a recombinant DNA molecule into their genomes. Preferred marker genes provide selective markers which confer resistance to a selective agent, such as an antibiotic or a herbicide. Any of the herbicides to which plants of this invention may be resistant are useful agents for selective markers. Potentially transformed cells are exposed to the selective agent. In the population of surviving cells will be those cells where, generally, the resistance-conferring gene is integrated and expressed at sufficient levels to permit cell survival. Cells may be tested further to confirm stable integration of the exogenous DNA. Commonly used selective marker genes include those conferring resistance to antibiotics such as kanamycin and paromomycin (nptII), hygromycin B (aph IV), spectinomycin (aadA) and gentamycin (aac3 and aacC4) or resistance to herbicides such as glufosinate (bar or pat), dicamba (DMO) and glyphosate (aroA or EPSPS). Examples of such selectable markers are illustrated in U.S. Pat. Nos. 5,550,318; 5,633,435; 5,780,708 and 6,118,047. Markers which provide an ability to visually screen transformants can also be employed, for example, a gene expressing a colored or fluorescent protein such as a luciferase or green fluorescent protein (GFP) or a gene expressing a beta-glucuronidase or uidA gene (GUS) for which various chromogenic substrates are known.

Plant cells that survive exposure to the selective agent, or plant cells that have been scored positive in a screening assay, may be cultured in regeneration media and allowed to mature into plants. Developing plantlets regenerated from transformed plant cells can be transferred to plant growth mix, and hardened off, for example, in an environmentally controlled chamber at about 85% relative humidity, 600 ppm CO₂, and 25-250 microeinsteins m⁻² s⁻¹ of light, prior to transfer to a greenhouse or growth chamber for maturation. Plants are regenerated from about 6 weeks to 10 months after a transformant is identified, depending on the initial tissue, and plant species. Plants may be pollinated using conventional plant breeding methods known to those of skill in the art and seed produced, for example self-pollination is commonly used with transgenic corn. The regenerated transformed plant or its progeny seed or plants can be tested for expression of the recombinant DNA and selected for the presence of enhanced agronomic trait.

Transgenic Plants and Seeds

Transgenic plants derived from transgenic plant cells having a transgenic nucleus of this invention are grown to generate transgenic plants having an enhanced trait as compared to a control plant and produce transgenic seed and haploid pollen of this invention. Such plants with enhanced traits are identified by selection of transformed plants or progeny seed for the enhanced trait. For efficiency a selection method is designed to evaluate multiple transgenic plants (events) comprising the recombinant DNA, for example multiple plants from 2 to 20 or more transgenic events. Transgenic plants grown from transgenic seed provided herein demonstrate improved agronomic traits that contribute to increased yield or other trait that provides increased plant value, including, for example, improved seed quality. Of particular interest are plants having enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil.

Table 1 provides a list of protein encoding DNA (“genes”) that are useful as recombinant DNA for production of transgenic plants with enhanced agronomic trait, the elements of Table 1 are described by reference to: “PEP SEQ ID NO” identifies an amino acid sequence from SEQ ID NO: 308 to 614. “NUC SEQ ID NO” identifies a DNA sequence from SEQ ID NO:1 to 307. “Gene ID” refers to an arbitrary identifier. “Gene Name” denotes a common name for the protein encoded by the recombinant DNA preceded by the abbreviated genus and species as fully defined in the sequence listing. The + or − preceding the gene name indicates whether the protein is expressed (+) or suppressed (−) in plants to provide an enhanced trait.

TABLE 1 NUC PEP SEQ SEQ ID ID NO NO Gene ID Gene Name 1 308 1174936:1 +Os.AlaAT 2 309 Mnom000960 −Gm.G1543 like 1 3 310 Mnom001103 +Gm.Hyh 4 311 1194741:1 −Gh.IDA-like 1:3:1 5 312 Mnom001209 +Zm.SCPL21 6 313 Mnom001263 +Zm.Tga1 7 314 Mnom001273 +Zm.A1ZM023275_at_Adenosine 5′-phosphosulfate reductase 8 8 315 Mnom001274 +Zm.A1ZM056190_at_B0806H07.1 protein 9 316 Mnom001282 −Zm.A1ZM009253_at_Hypothetical protein OJ1254_E07.2-1 10 317 1174117:1 +Ss.CGPG7432_hypothetical protein S||1109 11 318 1174129:1 +Xn.CGPG6757_Aspartate Aminotransferase 12 319 1169877:1 +Cg.D-lactate dehydrogenase (EC:1.1.1.28) 13 320 Mnom001350 +Ns.glycerol dehydrogenase (NADP+) (EC:1.1.1.72) 14 321 PHE0015383 +Bs.AldA (L-ALANINE DEHYDROGENASE) (EC:1.4.1.1) 15 322 Mnom001361 +Mt.Thiolase 16 323 Mnom001363 +Gh.Thiolase 17 324 Mnom001367 −Gm.Soul 18 325 Mnom001370 +Gm.X. bovienii spartate kinase_T359I 19 326 Mnom001372 +Gm.X.nematophila AK_T359I 20 327 Mnom001373 +Ec. metL AK-HSDS 21 328 1186474:1 +Gm.PIL-B 22 329 PHE0024821 +Gm.PIL-B 23 330 1184139:1 +At.PIF4 24 331 PHE0003348 +At.PIF4 25 332 1181794:1 +Gm.PICOT 26 333 1185540:1 +Ag.PRS 27 334 1184141:1 +At.CaMK4/At.CRK3 28 335 1183499:1 +At.GLN1; 1 29 336 1184141:1 +At.CaMK4/At.CRK3 30 337 1183676:1 +Gm.Myc:Dam 31 338 Mnom001401 +Gm.At.Hy5:Myc:Dam 32 339 1184600:1 +Gm.At.G1988:Myc:DAM 33 340 Mnom001436 +Rp.indole-3-pyruvate decarboxylase 34 341 Mnom001439 +At.tryptophane monooxygenase 35 342 Mnom001440 +Pf.tryptophane monooxygenase 36 343 Mnom001453 +Ta.Fd Nitrite reductase 37 344 1181413:1 −Zm.14-3-3 corn homolog (Chi) 38 345 Mnom001471 +Zm.PMH1 39 346 1193022:1 −Zm.NAC6-like 40 347 Mnom001521 +Gm.ERL2 41 348 1168947:1 +At.G1543-native-delta 1-21 42 349 1170457:1 +At.G1543 Kozak Variant (aa 1-4, changed MIKL to MASS) 43 350 PHE0001187 −Zm.spa1-like 44 351 Mnom001594 −Zm.SLAC1 45 352 1171516:1 +St.Pox1 46 353 PHE0022652 +St.Pox1 47 354 Mnom001612 +Zm.HOS15 48 355 Mnom001615 −Zm.ABI5 49 356 Mnom001618 +Le.SlGalLDH 50 357 Mnom001628 −Zm.NAC-like 51 358 Mnom001629 +Zm.NFD1 52 359 Mnom001636 +Zm.POZ domain protein 53 360 Mnom001641 −Zm.Putative nodule inception protein 54 361 Mnom001642 −Zm.ammonium transporter 55 362 Mnom001643 −Zm.POZ domain protein 56 363 Mnom001644 −Zm.isp4, oligopeptide transport 57 364 Mnom001645 −Zm.unknown 58 365 Mnom001647 −Zm.RAD-like 59 366 Mnom001651 +Pp.Formiminotransferase cyclodeaminase 60 367 Mnom001652 +Zm.Formiminotransferase cyclodeaminase 61 368 Mnom001653 +Cr.Na/glucose cotransporter 62 369 Mnom001661 +Cr.Cysteine sulfinate desulfinase/cysteine desulfurase, NifS 63 370 1194860:1 +Cr.Cysteine desulfurase (seleno-cysteine lyase) 64 371 Mnom001665 +Ne.ammonium monooxygenase subunit A 65 372 1180851:1 +PMEI1 66 373 Mnom001747 −Gm.MBD6 Soy homolog 67 374 1170440:1 +Gm.Col9 MRT3847_269613C 68 375 1174098:1 +GITI.PHD_SET 69 376 1183497:1 +Gm.G2512-like_AP2 domain transcriptional regulator 70 377 1188159:1 +Gm.G565-like_bZIP domain transcriptional regulator 71 378 1170356:1 +Gm.G2113-like_AP2 domain transcriptional regulator 72 379 1170348:1 +Gm.CGPG3920_expressed protein 73 380 1170352:1 +Gm.CGPG7836_ethylene responsive element binding factor_G1005-like 74 381 1170996:1 +At.Hsp101 75 382 1170342:1 +At.PEL1 76 383 1170354:1 +Gm.Unknown G1988 soybean heat induced gene 77 384 PHE0024001 −Zm.phosphoglucomutase 78 385 1190095:1 −Zm.phosphoglucomutase 79 386 Mnom001839 −Zm.ATHB16 HD-Zip I transcription factor 80 387 Mnom001840 −Zm.SOMNUS CCCH-type zinc finger protein 81 388 Mnom001841 −Zm.zinc finger protein 10 82 389 1174488:1 +Zm.basic helix-loop-helix (bHLH) family protein (AT5G50915-like) 83 390 Mnom001851 +At.Arginosuccinate synthase 84 391 Mnom001869 +Zm.Glutaredoxin domain containing protein 85 392 1175917:1 +Ec.AsnA 86 393 Mnom001877 −Zm.FNR 87 394 Mnom001880 +Ss.ntcA 88 395 Mnom001900 −Gm.FW2.2 like, PLAC8 89 396 Mnom001903 +Mt.MTP_Ac160241 90 397 Mnom001915 −Zm.MYB74 91 398 Mnom001921 −Zm.ASA1 92 399 Mnom001922 −Zm.WRKY72 93 400 Mnom001925 −Zm.CND41 homolog 94 401 Mnom001930 +Cr.Glutamine synthetase (GLN4) 95 402 Mnom001933 +Cr.Ferredoxin-dependent glutamate synthase (GSF1) 96 403 Mnom001936 +Cr.nitrite transporter NAR1 97 404 Mnom001937 −Zm.Initiator-binding protein; IBP2 98 405 1173274:1 +At.Rif1 (Resistant to inhibition by FSM) 99 406 1175406:1 +At.Glyoxylate Reductase 2 100 407 1178997:1 +At.Glyoxylate Reductase 1 101 408 1194803:1 +Cr.CPYC type (GRX1) 102 409 1194791:1 +Cr.CGFS type (GRX3) 103 410 1178100:1 +At.wt G1543 104 411 1184554:1 +At.G1543 (R138, 142A) 105 412 1178103:1 +At.G1543 NNO 106 413 1170936:1 −Zm.TAA1 107 414 Mnom001989 +Zm.Ghd7-2 108 415 Mnom001991 −Zm.MBD6 109 416 Mnom001992 −Zm.MBD10 110 417 1175891:1 +Zm.CGPG9187_Putative aurora-related kinase 111 418 1174476:1 +At.CGPG4567_Hypothetical protein AT4g11250 112 419 1181001:1 +At.CGPG3781_MADS-box protein-like 113 420 1158841:1 +Pd.PHE0017217_SCF5A8 114 421 1178370:1 +At.HDG11 115 422 1178370:1 +At.HDG11 116 423 PHE0024161 +At.HDG11 117 424 PHE0024161 +At.HDG11 118 425 Mnom002088 −Zm.Arginase (EC 3.5.3.1) 119 426 1195680:1 −Zm.KINB1 120 427 Mnom002104 +Gm.Nuclear factor Y NFYA5 121 428 Mnom002105 +Gm.Nuclear factor Y NFYA5 122 429 Mnom002106 +Gm.Nuclear factor Y NFYA5 123 430 Mnom002107 +Gm.Nuclear factor Y NFYA5 124 431 Mnom002108 +Gm.Nuclear factor Y NFYA5 125 432 Mnom002115 +At.phyB 126 433 1176524:1 +Gm.CGPG5320_HMG1/ 2-like protein 127 434 1183504:1 +Zm.SPRY 128 435 1183504:1 +Zm.SPRY 129 436 1180104:1 +Zm.RRM1 130 437 1180104:1 +Zm.RRM1 131 438 1182647:1 +Zm.RRM2 132 439 1182647:1 +Zm.RRM2 133 440 1182645:1 +Zm.RRM3 134 441 1182645:1 +Zm.RRM3 135 442 1182640:1 +Zm.KH2/ERA-like 136 443 1182640:1 +Zm.KH2/ERA-like 137 444 1182640:1 +Zm.KH2/ERA-like 138 445 1180839:1 +Os.BIRF1 (Benzathiodiazole induced ring finger protein 1) 139 446 PHE0024283 +Os.BIRF1 (Benzathiodiazole induced ring finger protein 1) 140 447 PHE0024283 +Os.BIRF1 (Benzathiodiazole induced ring finger protein 1) 141 448 1180839:1 +Os.BIRF1 (Benzathiodiazole induced ring finger protein 1) 142 449 Mnom002224 +Zm.CTR2 143 450 1188219:1 +Rs.glyoxylate reductase 144 451 1187645:1 +Bh.alanine dehydrogenase 145 452 1187645:1 +Bh.alanine dehydrogenase 146 453 1147250:1 +Zm.PHE0010151_glutaminyl-tRNA synthetase 147 454 PHE0024437 −Bn.MAX1 148 455 PHE0019867 +At.GLN1; 1 149 456 1185568:1 +At.HYH (HY5 Homolog) S24A mutant 150 457 PHE0024430 +At.HYH (HY5 Homolog) 151 458 1184541:1 +Ss.high-affinity branched-chain amino acid transport protein; BraE 152 459 1195300:1 +Ss.high-affinity branched-chain amino acid transport protein; BraE 153 460 1183470:1 +Ss.aspartate 1-decarboxylase 154 461 1183502:1 +Ss.Bifunctional imidazoleglycerol- phosphate dehydratase; histidinol-phosphatase 155 462 1184961:1 +Ss.Glutamate permease 156 463 PHE0024436 +Ss.Glutamate permease 157 464 1184550:1 +Ss.glutamine ABC transporter 158 465 PHE0024437 +Ss.glutamine ABC transporter 159 466 Mnom002329 +Ns.Nitrite/Nitrate permease 160 467 1183533:1 +Ss.alanine dehydrogenase 161 468 1181792:1 +Cg.PHE0007792_nitroreductase 162 469 Mnom002337 +Zm.Putative acid phosphatase 163 470 1183509:1 +At.HYH (HY5 Homolog) 164 471 1194207:1 +Bn.SUM2 165 472 1184951:1 +At.da1-1 166 473 1184947:1 +Zm.da1-1 167 474 Mnom002354 −Zm.BB 168 475 Mnom002372 +Zm.Putative UVB-resistance protein UVR8 169 476 Mnom002381 −Zm.Putative sulfate transporter 170 477 Mnom002382 −Zm.Putative UVB-resistance protein UVR8 171 478 Mnom002383 −Zm.Putative transfactor 172 479 Mnom002384 −Zm.Putative trehalose-6- phosphate synthase 173 480 Mnom002385 −Zm.Putative polypyrimidine tract- binding protein homolog 174 481 Mnom002387 −Zm.Putative aspartic proteinase nepenthesin I 175 482 Mnom002388 −Zm.Os08g0542700 protein 176 483 Mnom002389 −Zm.unknown protein 177 484 Mnom002390 −Zm.Dual-specificity protein-like phosphatase 3 178 485 Mnom002391 −Zm.Hypothetical protein 179 486 Mnom002404 −Zm.PUB22/23-like protein1 180 487 Mnom002405 −Zm.PUB22/23-like protein2 181 488 Mnom002408 +At.CLV3 182 489 Mnom002410 +At.mCLV3 183 490 1183506:1 +At.CLE-like AT5G59305.1 precursor 184 491 1183506:1 +At.CLE-like AT5G59305.1 precursor 185 492 Mnom002414 +At.CLE-like AT5G59305.1 mature 186 493 Mnom002426 +Gh.Skd1 (DQ444282) 187 494 1163080:1 +At.ERF1 188 495 1179434:1 +Zm.homolog Basic Transcription Factor 2 (CGPG1840) 189 496 Mnom002481 +Zm.ATH1-1 190 497 1187102:1 +Zm.ATH1-2 191 498 1186139:1 +Zm.ZRP4 192 499 Mnom002490 +Zm.LSH1 193 500 Mnom002494 +Zm.Putative cytochrome P450 194 501 1183523:1 +At.NRT1.5 (nitrate transporter 1.5) 195 502 Mnom002497 −Zm.homolog of CIPK8 (Calcineurin B-like (CBL)- interacting protein kinase 8) 196 503 1184967:1 +Zm.RINUE8 197 504 1188565:1 +At.phyA 198 505 Mnom002520 +At.phyA Y242H 199 506 1184536:1 +At.phyA F389A 200 507 PHE0024283 +Os.BIRF1 (Benzathiodiazole induced ring finger protein 1) 201 508 PHE0024283 +Os.BIRF1 (Benzathiodiazole induced ring finger protein 1) 202 509 1181806:1 +Ec. cspG - D63344 203 510 1181808:1 +At.PHE0007559_OXI1 (OXIDATIVE SIGNAL- INDUCIBLE1) 204 511 1183495:1 +Zm.PHE0007208_Putative polysialic acid capsule expression protein 205 512 PHE0007541 +Zm.PHE0007541_A1ZM008639_at 206 513 1181787:1 +Dh.PHE0007554_BCCT transporter 207 514 1181764:1 +Ta.PHE0007755_Td25a 208 515 1181792:1 +Cg.PHE0007792_nitroreductase 209 516 1181783:1 +At.PHE0009197_PF02519- Auxin_inducible-9 210 517 1184534:1 +At.TZP, At5g43630 211 518 Mnom002555 −Gm.PUB22/PUB23 like 212 519 1181848:1 +Os.bZIP23(Basic leucine zipper 23) 213 520 1185549:1 +At.ZEP (Zeaxanthin epoxidase) 214 521 1184133:1 +Gm.Transcription factor LIM 215 522 1181771:1 +Gm.Putative pollen specific protein 216 523 Mnom002564 +Gm.unknown protein contain domains similar to protein serine/threonine kinase 217 524 Mnom002565 +Gm.MADS-box protein 218 525 1181802:1 +Gm.Seven in absentia protein family putative 219 526 1183468:1 +At.bHLH047 220 527 Mnom002582 +At.MYB114 221 528 Mnom002603 +At.G2930 222 529 Mnom002604 +At.G2784 223 530 1087245:1 +At.G154 MADS-box protein 224 531 Mnom002645 +At.Twin Sister of FT (TSF) protein 225 532 Mnom002652 −Zm.homolog of Arabidopsis peptide transporter 5(PTR5) 226 533 Mnom002653 +At.Transport protein associated with antigen processing2 (TAP2)/ Aluminium sensitive1 (ALS1) 227 534 Mnom002662 +Cc.Asparagine synthetase codon optimized 228 535 Mnom002667 +Cr.Acetate kinase 229 536 Mnom002668 +Cr.phosphate acetyltransferase 230 537 Mnom002669 +Cr.Lysine decarboxylase 231 538 1066582:1 +At.Aromatic and neutral amino acid transporter 1 (ANT1) 232 539 PHE0009168 +Le.PHE0009168_JERF3 (Jasmonate and ethylene responsive factor 3) 233 540 Mnom002676 +Gm.CHI 234 541 Mnom002706 −Zm.RAA1 235 542 Mnom002708 +Zm.Albino3 236 543 1186548:1 +At.LPA2 (Low PSII Accumulation2) 237 544 Mnom002714 −Zm.Lew1 (leaf wilting1) 238 545 Mnom002716 −Zm.eIF-5A 239 546 1190083:1 +Vf.Amino acid permease 1 (AAP1) 240 547 Mnom002721 +Gm.G1481 a homolog of AtG1481 241 548 PHE0024832 +At.Nitrate Transporter 1.5 242 549 1190115:1 +Pp.HB4 243 550 1187139:1 +At.HB1 244 551 1187139:1 +At.HB1 245 552 1187139:1 +At.HB1 246 553 1188214:1 +Zm.PP2C 247 554 1189232:1 +Zm.PP2C 248 555 Mnom002750 +Zm.PP2C 249 556 1188212:1 +Os.PP2C 250 557 PHE0006743 +At.CGPG151 putative two-component response regulator protein 251 558 PHE0004397 +Os.G3848 252 559 PHE0006386 +At.CGPG1137 expressed protein 253 560 1187105:1 +Zm.tubby 4 254 561 Mnom002784 +At.Cationic amino acid transporter1 (CAT1) 255 562 1194701:1 +At.Cationic amino acid transporter6 (CAT6) 256 563 1194425:1 +Cr.fructose 6 phosphate aldolase 257 564 PHE0025040 +Cr.Acetate kinase 258 565 PHE0025042 +Cr.Lysine decarboxylase 259 566 1190117:1 +At.galactose-binding lectin family protein 260 567 1190127:1 +At.zinc finger (C3HC4- type RING finger) family protein 261 568 1191292:1 +At.putative amino acid transport protein 262 569 1190123:1 +At.zinc finger (C3HC4- type RING finger) family protein 263 570 1194201:1 +At.lesion inducing protein-related 264 571 PHE0025430 +Ta.DNA binding with one finger (TaDOF1) 265 572 1187623:1 +Ta.DNA binding with one finger (TaDOF1) 266 573 1191310:1 +Ta.DNA binding with one finger (TaDOF1) 267 574 Mnom002849 +Bn.G1988 homologue 268 575 PHE0025389 +At.phyA S602A 269 576 1192540:1 +At.phyA S602A 270 577 1192540:1 +At.phyA S602A 271 578 Mnom002860 +At.ammonium transporter At Amt1; 5 272 579 Mnom002864 +At.TF At5g56860 273 580 Mnom002882 +At.Gene 5160 274 581 Mnom002886 +At.RUS1 275 582 Mnom002887 +Sl.TERF1 (Tomato ethylene response factor 1) 276 583 1195225:1 +Ca.Pathogen-induced membrane protein 1 277 584 Mnom002889 +Zm.DEK1 (Defective Kernel 1) 278 585 Mnom002891 +Gm.BiPD(Endoplasmic reticulum HSC70-cognate binding protein) 279 586 1191787:1 +Ta.W55a 280 587 1194731:1 +At.WRKY25 281 588 Mnom002895 +At.WRKY33 282 589 Mnom002896 +Ps.Sym37 283 590 Mnom002898 +Mt.DMI2 284 591 Mnom002899 +Pv.NAS2 (asparagines synthetase 2) 285 592 1187621:1 +Os.Gene 179_MonNom 286 593 1187788:1 +Vv.Gene 197 287 594 1194056:1 +Os.CGFS type glutaredoxin (mitochondrial form) 288 595 Mnom002995 +Os.Multidomian Monothiol glutaredoxin 289 596 1194054:1 +At.Monothiol glutaredoxin 290 597 Mnom003001 +At.GTPase ObgE family 291 598 Mnom003035 +At.RGE1(RETARDED GROWTH OF EMBRYO1) 292 599 1192245:1 +Zm.Hypothetical protein specific to corn endosperm 293 600 1194063:1 +At.leucine-rich repeat family protein 294 601 PHE0021991 +At.1g03190 (AtXPD) 295 602 1193217:1 +At.CoAse3(Coenzyme A pyrophosphatase 3) 296 603 Mnom003125 +At.MRB1 (Membrane Related Bigger 1) 297 604 1194048:1 +Ca.RZFP1 (RING Zinc Finger Protein 1) 298 605 Mnom003151 +Gm.G154 homologue3 299 606 Mnom003152 +Gh.G154 homologue 300 607 Mnom003153 +Gm.AT1G47670 - LYS/HIS TRANSPORTER 7 301 608 Mnom003154 +Gm.AT5G23810 - AAP7 302 609 Mnom003157 +Gm.GXGXG domain containing protein 303 610 Mnom003158 +Gm.DUF_domain containing protein 304 611 Mnom003159 +Gm.AT5G65660 - hydroxyproline-rich glycoprotein family protein 305 612 Mnom003178 +Le.AIM1(Abscisic acid- induced myb1) 306 613 Mnom003189 +Ta.Wheat hemoglobin HMP reductase fusion 307 614 Mnom003190 +Pp.Flavohemoglobin (codon optimized) Selection Methods for Transgenic Plants with Enhanced Agronomic Trait

Within a population of transgenic plants each regenerated from a plant cell having a nucleus with recombinant DNA many plants that survive to fertile transgenic plants that produce seeds and progeny plants will not exhibit an enhanced agronomic trait. Selection from the population is necessary to identify one or more transgenic plant cells having a transgenic nucleus that can provide plants with the enhanced trait. Transgenic plants having enhanced traits are selected from populations of plants regenerated or derived from plant cells transformed as described herein by evaluating the plants in a variety of assays to detect an enhanced trait, e.g. enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil. These assays also may take many forms including, but not limited to, direct screening for the trait in a greenhouse or field trial or by screening for a surrogate trait. Such analyses can be directed to detecting changes in the chemical composition, biomass, physiological properties, morphology of the plant. Changes in chemical compositions such as nutritional composition of grain can be detected by analysis of the seed composition and content of protein, free amino acids, oil, free fatty acids, starch or tocopherols. Changes in biomass characteristics can be made on greenhouse or field grown plants and can include plant height, stem diameter, root and shoot dry weights; and, for corn plants, ear length and diameter. Changes in physiological properties can be identified by evaluating responses to stress conditions, for example assays using imposed stress conditions such as water deficit, nitrogen deficiency, cold growing conditions, pathogen or insect attack or light deficiency, or increased plant density. Changes in morphology can be measured by visual observation of tendency of a transformed plant with an enhanced agronomic trait to also appear to be a normal plant as compared to changes toward bushy, taller, thicker, narrower leaves, striped leaves, knotted trait, chlorosis, albino, anthocyanin production, or altered tassels, ears or roots. Other selection properties include days to pollen shed, days to silking, leaf extension rate, chlorophyll content, leaf temperature, stand, seedling vigor, internode length, plant height, leaf number, leaf area, tittering, brace roots, stay green, stalk lodging, root lodging, plant health, barreness/prolificacy, green snap, and pest resistance. In addition, phenotypic characteristics of harvested grain may be evaluated, including number of kernels per row on the ear, number of rows of kernels on the ear, kernel abortion, kernel weight, kernel size, kernel density and physical grain quality.

Assays for screening for a desired trait are readily designed by those practicing in the art. The following illustrates useful screening assays for corn traits using hybrid corn plants. The assays can be readily adapted for screening other plants such as canola, cotton and soybean either as hybrids or inbreds.

Transgenic corn plants having nitrogen use efficiency are identified by screening in fields with three levels of nitrogen (N) fertilizer being applied, e.g. low level (0 N), medium level (80 lb/ac) and high level (180 lb/ac). Plants with enhanced nitrogen use efficiency provide higher yield as compared to control plants.

Transgenic corn plants having enhanced yield are identified by screening using progeny of the transgenic plants over multiple locations with plants grown under optimal production management practices and maximum weed and pest control. A useful target for improved yield is a 5% to 10% increase in yield as compared to yield produced by plants grown from seed for a control plant. Selection methods may be applied in multiple and diverse geographic locations, for example up to 16 or more locations, over one or more planting seasons, for example at least two planting seasons, to statistically distinguish yield improvement from natural environmental effects.

Transgenic corn plants having enhanced water use efficiency are identified by screening plants in an assay where water is withheld for a period to induce stress followed by watering to revive the plants. For example, a useful selection process imposes 3 drought/re-water cycles on plants over a total period of 15 days after an initial stress free growth period of 11 days. Each cycle consists of 5 days, with no water being applied for the first four days and a water quenching on the 5th day of the cycle. The primary phenotypes analyzed by the selection method are the changes in plant growth rate as determined by height and biomass during a vegetative drought treatment.

Transgenic corn plants having enhanced cold tolerance are identified by screening plants in a cold germination assay and/or a cold tolerance field trial. In a cold germination assay trays of transgenic and control seeds are placed in a growth chamber at 9.7° C. for 24 days (no light). Seeds having higher germination rates as compared to the control are identified as having enhanced cold tolerance. In a cold tolerance field trial plants with enhanced cold tolerance are identified from field planting at an earlier date than conventional Spring planting for the field location. For example, seeds are planted into the ground around two weeks before local farmers begin to plant corn so that a significant cold stress is exerted onto the crop, named as cold treatment. Seeds also are planted under local optimal planting conditions such that the crop has little or no exposure to cold condition, named as normal treatment. At each location, seeds are planted under both cold and normal conditions preferably with multiple repetitions per treatment.

Transgenic corn plants having seeds with increased protein and/or oil levels are identified by analyzing progeny seed for protein and/or oil. Near-infrared transmittance spectrometry is a non-destructive, high-throughput method that is useful to determine the composition of a bulk seed sample for properties listed in table 2.

TABLE 2 Typical sample(s): Whole grain corn and soybean seeds Typical analytical range: Corn - moisture 5-15%, oil 5-20%, protein 5-30%, starch 50-75%, and density 1.0-1.3%. Soybean - moisture 5-15%, oil 15-25%, and protein 35-50%.

Although the plant cells and methods of this invention can be applied to any plant cell, plant, seed or pollen, e.g. any fruit, vegetable, grass, tree or ornamental plant, the various aspects of the invention are preferably applied to corn, soybean, cotton, canola, alfalfa, wheat, rice, sugarcane, and sugar beet plants. In many cases the invention is applied to corn plants that are inherently resistant to disease from the Mal de R10Cuarto virus or the Puccina sorghi fungus or both.

The following examples are included to demonstrate aspects of the invention, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific aspects which are disclosed and still obtain a like or similar results without departing from the spirit and scope of the invention.

Example 1 Plant Expression Constructs

This example illustrates the construction of plasmids for transferring recombinant DNA into a plant cell nucleus that can be regenerated into transgenic plants.

A. Plant Expression Constructs for Corn Transformation

A base corn transformation vector pMON93039, as set forth in SEQ ID NO:36443, illustrated in Table 3, is fabricated for use in preparing recombinant DNA for Agrobacterium-mediated transformation into corn tissue.

TABLE 3 Coordinates of Function Name Annotation SEQ ID NO: 36443 Agrobacterium B-AGRtu.right border Agro right border sequence, 11364-11720 T-DNA transfer essential for transfer of T-DNA. Gene of interest E-Os.Act1 Upstream promoter region of the  19-775 expression rice actin 1 gene cassette E-CaMV.35S.2xA1- Duplicated35S A1-B3 domain  788-1120 B3 without TATA box P-Os.Act1 Promoter region of the rice actin 1 1125-1204 gene L-Ta.Lhcb1 5′ untranslated leader of wheat 1210-1270 major chlorophyll a/b binding protein I-Os.Act1 First intron and flanking UTR exon 1287-1766 sequences from the rice actin 1 gene T-St.Pis4 3′ non-translated region of the 1838-2780 potato proteinase inhibitor II gene which functions to direct polyadenylation of the mRNA Plant selectable P-Os.Act1 Promoter from the rice actin 1 gene 2830-3670 marker L-Os.Act1 First exon of the rice actin 1 gene 3671-3750 expression I-Os.Act1 First intron and flanking UTR exon 3751-4228 cassette sequences from the rice actin 1 gene TS-At.ShkG-CTP2 Transit peptide region of 4238-4465 Arabidopsis EPSPS CR-AGRtu.aroA- Coding region for bacterial strain 4466-5833 CP4.nat CP4 native aroA gene. T-AGRtu.nos A 3′ non-translated region of the 5849-6101 nopaline synthase gene of Agrobacterium tumefaciens Ti plasmid which functions to direct polyadenylation of the mRNA. Agrobacterium B-AGRtu.left border Agro left border sequence, essential 6168-6609 T-DNA transfer for transfer of T-DNA. Maintenance in OR-Ec.oriV-RK2 The vegetative origin of replication 6696-7092 E. coli from plasmid RK2. CR-Ec.rop Coding region for repressor of 8601-8792 primer from the ColE1 plasmid. Expression of this gene product interferes with primer binding at the origin of replication, keeping plasmid copy number low. OR-Ec.ori-ColE1 The minimal origin of replication 9220-9808 from the E. coli plasmid ColE1. P-Ec.aadA-SPC/STR Promoter for Tn7 10339-10380 adenylyltransferase (AAD(3″)) CR-Ec.aadA- Coding region for Tn7 10381-11169 SPC/STR adenylyltransferase (AAD(3″)) conferring spectinomycin and streptomycin resistance. T-Ec.aadA-SPC/STR 3′ UTR from the Tn7 11170-11227 adenylyltransferase (AAD(3″)) gene of E. coli.

To construct transformation vectors for expressing a protein identified in Table 1, primers for PCR amplification of the protein coding nucleotides are designed at or near the start and stop codons of the coding sequence, in order to eliminate most of the 5′ and 3′ untranslated regions. The protein coding nucleotides are inserted into the base vector in the gene of interest expression cassette at an insertion site, i.e. between the intron element (coordinates 1287-1766) and the polyadenylation element (coordinates 1838-2780).

To construct transformation vectors for suppressing a protein identified in Table 1, the amplified protein coding nucleotides are assembled in a sense and antisense arrangement and inserted into the base vector at the insertion site in the gene of interest expression cassette to provide transcribed RNA that will form a double-stranded RNA for RNA interference suppression of the protein. More specifically, the sense and anti-sense DNA is derived from an endogenous corn gene that expresses a corn protein with an amino acid sequence of SEQ ID NO: 316, 344, 346, 350-351, 355, 357, 360-365, 384-388, 393, 397-400, 404, 413, 415-416, 425-426, 474, 476-487, 502, 532, 541, or 544-545 or the corn homolog of SEQ ID NOs:309, 311, 324, 373, 395, 454, or 518.

B. Plant Expression Constructs for Soy and Canola Transformation

Vectors for use in transformation of soybean and canola tissue are prepared having the elements of expression vector pMON82053 (SEQ ID NO: 36444) as shown in Table 4 below.

TABLE 4 Coordinates of Function Name Annotation SEQ ID NO: 36444 Agrobacterium T- B-AGRtu.left border Agro left border sequence, essential for 6144-6585 DNA transfer transfer of T-DNA. Plant selectable P-At.Act7 Promoter from the Arabidopsis actin 7 gene 6624-7861 marker expression L-At.Act7 5′UTR of Arabidopsis Act7 gene cassette I-At.Act7 Intron from the Arabidopsis actin7 gene TS-At.ShkG-CTP2 Transit peptide region of Arabidopsis 7864-8091 EPSPS CR-AGRtu.aroA- Synthetic CP4 coding region with dicot 8092-9459 CP4.nno_At preferred codon usage. T-AGRtu.nos A 3′ non-translated region of the nopaline 9466-9718 synthase gene of Agrobacterium tumefaciens Ti plasmid which functions to direct polyadenylation of the mRNA. Gene of interest P-CaMV.35S-enh Promoter for 35S RNA from CaMV  1-613 expression cassette containing a duplication of the −90 to −350 region. T-Gb.E6-3b 3′ untranslated region from the fiber protein  688-1002 E6 gene of sea-island cotton. Agrobacterium T- B-AGRtu.right Agro right border sequence, essential for 1033-1389 DNA transfer border transfer of T-DNA. Maintenance in E. coli OR-Ec.oriV-RK2 The vegetative origin of replication from 5661-6057 plasmid RK2. CR-Ec.rop Coding region for repressor of primer from 3961-4152 the ColE1 plasmid. Expression of this gene product interferes with primer binding at the origin of replication, keeping plasmid copy number low. OR-Ec.ori-ColE1 The minimal origin of replication from the 2945-3533 E. coli plasmid ColE1. P-Ec.aadA-SPC/STR Promoter for Tn7 adenylyltransferase 2373-2414 (AAD(3″)) CR-Ec.aadA- Coding region for Tn7 adenylyltransferase 1584-2372 SPC/STR (AAD(3″)) conferring spectinomycin and streptomycin resistance. T-Ec.aadA-SPC/STR 3′ UTR from the Tn7 adenylyltransferase 1526-1583 (AAD(3″)) gene of E. coli.

To construct transformation vectors for expressing a protein identified in Table 1, primers for PCR amplification of the protein coding nucleotides are designed at or near the start and stop codons of the coding sequence, in order to eliminate most of the 5′ and 3′ untranslated regions. The protein coding nucleotides are inserted into the base vector in the gene of interest expression cassette at an insertion site, i.e. between the promoter element (coordinates 1-613) and the polyadenylation element (coordinates 688-1002).

To construct transformation vectors for suppressing a protein identified in Table 1, the amplified protein coding nucleotides are assembled in a sense and antisense arrangement and inserted into the base vector at the insertion site in the gene of interest expression cassette to provide transcribed RNA that will form a double-stranded RNA for RNA interference suppression of the protein. More specifically, for soybean the sense and anti-sense DNA is derived from an endogenous soybean gene that expresses a soybean protein with an amino acid sequence of SEQ ID NOs: 309, 324, 373, 395, 518 or is a soybean homolog of SEQ ID NOs: 311, 316, 344, 346, 350-351, 355, 357, 360-365, 384-388, 393, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 532, 541, or 544-545, and for canola the sense and anti-sense DNA is derived from an endogenous canola gene that encodes the canola homolog of SEQ ID NOs: 309, 311, 316, 324, 344, 346, 350-351, 355, 357, 360-365, 373, 384-388, 393, 395, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 518, 532, 541, or 544-545.

C. Cotton Transformation Vector

Plasmids for use in transformation of cotton tissue are prepared with elements of expression vector pMON99053 (SEQ ID NO: 36445) as shown in Table 5 below.

TABLE 5 Coordinates of SEQ ID NO: Function Name Annotation 36445 Agrobacterium T- B-AGRtu.right border Agro right border sequence, essential for  1-357 DNA transfer transfer of T-DNA. Gene of interest Exp-CaMV.35S- Enhanced version of the 35S RNA  388-1091 expression enh + Ph.DnaK promoter from CaMV plus the petunia cassette hsp70 5′ untranslated region T-Ps.RbcS2-E9 The 3′ non-translated region of the pea 1165-1797 RbcS2 gene which functions to direct polyadenylation of the mRNA. Plant selectable Exp-CaMV.35S Promoter and 5′ untranslated region from 1828-2151 marker expression the 35S RNA of CaMV cassette CR-Ec.nptII-Tn5 Coding region for neomycin 2185-2979 phosphotransferase gene from transposon Tn5 which confers resistance to neomycin and kanamycin. T-AGRtu.nos A 3′ non-translated region of the nopaline 3011-3263 synthase gene of Agrobacterium tumefaciens Ti plasmid which functions to direct polyadenylation of the mRNA. Agrobacterium T- B-AGRtu.left border Agro left border sequence, essential for 3309-3750 DNA transfer transfer of T-DNA. Maintenance in E. coli OR-Ec.oriV-RK2 The vegetative origin of replication from 3837-4233 plasmid RK2. CR-Ec.rop Coding region for repressor of primer from 5742-5933 the ColE1 plasmid. Expression of this gene product interferes with primer binding at the origin of replication, keeping plasmid copy number low. OR-Ec.ori-ColE1 The minimal origin of replication from the 6361-6949 E. coli plasmid ColE1. P-Ec.aadA-SPC/STR Promoter for Tn7 adenylyltransferase 7480-7521 (AAD(3″)) CR-Ec.aadA-SPC/STR Coding region for Tn7 adenylyltransferase 7522-8310 (AAD(3″)) conferring spectinomycin and streptomycin resistance. T-Ec.aadA-SPC/STR 3′ UTR from the Tn7 adenylyltransferase 8311-8368 (AAD(3″)) gene of E. coli.

To construct transformation vectors for expressing a protein identified in Table 1, primers for PCR amplification of the protein coding nucleotides are designed at or near the start and stop codons of the coding sequence, in order to eliminate most of the 5′ and 3′ untranslated regions. The protein coding nucleotides are inserted into the base vector in the gene of interest expression cassette at an insertion site, i.e. between the promoter element (coordinates 388-1091) and the polyadenylation element (coordinates 1165-1797).

To construct transformation vectors for suppressing a protein identified in Table 1, the amplified protein coding nucleotides are assembled in a sense and antisense arrangement and inserted into the base vector at the insertion site in the gene of interest expression cassette to provide transcribed RNA that will form a double-stranded RNA for RNA interference suppression of the protein. More specifically, the sense and anti-sense DNA is derived from an endogenous cotton gene that encodes the cotton homolog of SEQ ID NO: 309, 311, 316, 324, 344, 346, 350-351, 355, 357, 360-365, 373, 384-388, 393, 395, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 518, 532, 541, or 544-545.

D. Plant Expression Constructs for Gene Stacking in Corn.

A base corn transformation vector pMON96782, as set forth in SEQ ID NO: 36446, illustrated in Table 6, is fabricated for use in preparing recombinant DNA for Agrobacterium-mediated transformation into corn tissue.

TABLE 6 Coordinates of Function Name Annotation SEQ ID NO: 36446 Agrobacterium B-AGRtu.right border Agro right border sequence,  1-357 T-DNA transfer essential for transfer of T-DNA. Gene of interest P-Os.Act1 Promoter region of the rice actin 1  403-1243 expression gene cassette 1 L-Os.Act1 5′ untranslated leader of rice actin 1 1244-1323 gene I-Os.Act1 First intron and flanking UTR exon 1324-1801 sequences from the rice actin 1 gene T-Ta.Hsp17 3′ un-translated region of wheat low 1834-2043 molecular weight heat shock protein gene Gene of interest E-Os.Act1 Upstream Promoter region of rice 2136-2892 expression actin 1 cassette 2 E-CaMV.35S.2xA1- 35S A1-B3 Domain 2905-2937 B3 P-Os.Act1 Promoter from rice actin gene 3242-3321 L-Ta.Lhcb1 5′ untranslated leader from wheat 3327-3387 chlorophyll protein I-Os.Act1 Intron and 5′ untranslated region 3404-3883 from rice actin 1 gene T-AGRtu.tr7 3′ untranslated region from 3918-4425 “transcript 7” of Agrobacterium Plant selectable P.Os.TubA Promoter of alpha-tubulin gene of 4452-5650 marker rice expression L.Os.TubA 5′ untranslated region of an alpha 5651-5736 cassette tubulin from rice I.Os.TubA Intron 1 of an alpha tubulin from 5737-6632 rice. Ts.Ta.waxy.nno_Zm Chloroplast transit peptide from 6637-6846 wheat starch synthase Cr.AGRtu.aroA- CP4 EPSPS gene 6847-8214 CP4.nno_Zm T.Os.TubA 3′ untranslated region of alpha 8219-8800 tubulin from rice Agrobacterium B-AGRtu.left border Left border sequence for T-DNA 8828-9269 T-DNA transfer transfer Maintenance in OR-Ec.oriV-RK2 Origin of replication from the E. coli 9356-9752 E. coli plasmid RK2. Cr-Ec.rop Coding region for repressor of 11261-11452 primer from ColE1 plasmid OR-Ec.ori-ColE1 Minimum origin of replication from 11880-12468 E. coli colE1 plasmid. P-Ec.aadA-SPC/STR Promoter for Tn7 12999-13040 adenylyltransferase gene CR-Ec.aadA- Coding region for Tn7 13041-13829 SPC/STR adenylyltransferase gene T-Ec.aadA-SPC/STR 3′ untranslated region from Tn7 13830-13887 adenylyltransferase gene

Primers for PCR amplification of protein coding nucleotides of the genes of interest are designed at or near the start and stop codons of the coding sequence, in order to eliminate most of the 5′ and 3′ untranslated regions. Protein coding regions of genes encoding a first and second protein of interest are amplified. The amplified region from the first gene of interest is cloned between nucleotides 1801 and 1834 of the base vector and the amplified region from the second gene of interest is cloned between nucleotides 3883 and 3918 of the base vector.

Example 2 Corn Transformation

This example illustrates transformation methods useful in producing a transgenic nucleus in a corn plant cell, and the plants, seeds and pollen produced from a transgenic cell with such a nucleus having an enhanced trait, i.e. enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil. A plasmid vector is prepared by cloning the DNA of SEQ ID NO:1 into the gene of interest expression cassette in the base vector for use in corn transformation of corn tissue provided in Example 1, Table 3.

For Agrobacterium-mediated transformation of corn embryo cells corn plants of a readily transformable line are grown in the greenhouse and ears are harvested when the embryos are 1.5 to 2.0 mm in length. Ears are surface sterilized by spraying or soaking the ears in 80% ethanol, followed by air drying Immature embryos are isolated from individual kernels on surface sterilized ears. Prior to inoculation of maize cells, Agrobacterium cells are grown overnight at room temperature Immature maize embryo cells are inoculated with Agrobacterium shortly after excision, and incubated at room temperature with Agrobacterium for 5-20 minutes. Immature embryo plant cells are then co-cultured with Agrobacterium for 1 to 3 days at 23° C. in the dark. Co-cultured embryos are transferred to selection media and cultured for approximately two weeks to allow embryogenic callus to develop. Embryogenic callus is transferred to culture medium containing 100 mg/L paromomycin and subcultured at about two week intervals. Transformed plant cells are recovered 6 to 8 weeks after initiation of selection.

For Agrobacterium-mediated transformation of maize callus immature embryos are cultured for approximately 8-21 days after excision to allow callus to develop. Callus is then incubated for about 30 minutes at room temperature with the Agrobacterium suspension, followed by removal of the liquid by aspiration. The callus and Agrobacterium are co-cultured without selection for 3-6 days followed by selection on paromomycin for approximately 6 weeks, with biweekly transfers to fresh media. Paromomycin resistant calli are identified about 6-8 weeks after initiation of selection.

To regenerate transgenic corn plants a callus of transgenic plant cells resulting from transformation and selection is placed on media to initiate shoot development into plantlets which are transferred to potting soil for initial growth in a growth chamber at 26° C. followed by a mist bench before transplanting to 5 inch pots where plants are grown to maturity. The regenerated plants are self-fertilized and seed is harvested for use in one or more methods to select seeds, seedlings or progeny second generation transgenic plants (R2 plants) or hybrids, e.g. by selecting transgenic plants exhibiting an enhanced trait as compared to a control plant.

The above process is repeated to produce multiple events of transgenic corn plant cells that are transformed with recombinant DNA from each of the genes identified in Table 1. Events are designed to produce in the transgenic cells one of the proteins identified in Table 1, except the proteins of SEQ ID NOs: 316, 344, 346, 350-351, 355, 357, 360-365, 384-388, 393, 397-400, 404, 413, 415-416, 425-426, 474, 476-487, 502, 532, 541, and 544-545, and the corn homolog of SEQ ID NOs: 309, 311, 324, 373, 395, 454, and 518 which are suppressed. Progeny transgenic plants and seed of the transformed plant cells are screened for enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil. From each group of multiple events of transgenic plants with a specific recombinant DNA from Table 1 the event that produces the greatest enhancement in yield, water use efficiency, nitrogen use efficiency, enhanced cold tolerance, enhanced seed protein and enhanced seed oil is identified and progeny seed is selected for commercial development.

Example 3 Soybean Transformation

This example illustrates plant transformation useful in producing a transgenic nucleus in a soybean plant cell, and the plants, seeds and pollen produced from a transgenic cell with such a nucleus having an enhanced trait, i.e. enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil.

For Agrobacterium mediated transformation, soybean seeds are imbided overnight and the meristem explants excised. The explants are placed in a wounding vessel. Soybean explants and induced Agrobacterium cells from a strain containing plasmid DNA with the gene of interest cassette and a plant selectable marker cassette are mixed no later than 14 hours from the time of initiation of seed imbibition, and wounded using sonication. Following wounding, explants are placed in co-culture for 2-5 days at which point they are transferred to selection media for 6-8 weeks to allow selection and growth of transgenic shoots. Resistant shoots are harvested approximately 6-8 weeks and placed into selective rooting media for 2-3 weeks. Shoots producing roots are transferred to the greenhouse and potted in soil. Shoots that remain healthy on selection, but do not produce roots are transferred to non-selective rooting media for an additional two weeks. Roots from any shoots that produce roots off selection are tested for expression of the plant selectable marker before they are transferred to the greenhouse and potted in soil.

The above process is repeated to produce multiple events of transgenic soybean plant cells that are transformed with recombinant DNA from each of the genes identified in Table 1. Events are designed to produce in the transgenic cells one of the proteins identified in Table 1, except the proteins of SEQ ID NOs: 309, 324, 373, 395, and 518 and the soybean homologs of SEQ ID NOs: 311, 316, 344, 346, 350-351, 355, 357, 360-365, 384-388, 393, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 532, 541, and 544-545, which are suppressed. Progeny transgenic plants and seed of the transformed plant cells are screened for enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced seed protein and enhanced seed oil. From each group of multiple events of transgenic plants with a specific recombinant DNA from Table 1 the event that produces the greatest enhancement in yield, water use efficiency, nitrogen use efficiency, enhanced cold tolerance, enhanced seed protein and enhanced seed oil is identified and progeny seed is selected for commercial development.

Example 4 Cotton Transgenic Plants with Enhanced Agronomic Traits

This example illustrates plant transformation useful in producing a transgenic nucleus in a cotton plant cell, and the plants, seeds and pollen produced from a transgenic cell with such a nucleus having an enhanced trait, i.e. enhanced water use efficiency, increased yield, enhanced nitrogen use efficiency and enhanced seed oil.

Transgenic cotton plants containing each recombinant DNA having a sequence of SEQ ID NO: 1 through SEQ ID NO: 307 are obtained by transforming with recombinant DNA from each of the genes identified in Table 1 using Agrobacterium-mediated transformation. The above process is repeated to produce multiple events of transgenic cotton plant cells that are transformed with recombinant DNA from each of the genes identified in Table 1. Events are designed to produce in the transgenic cells one of the proteins identified in Table 1, except the cotton homologs of the proteins of SEQ ID NOs: 309, 311, 316, 324, 344, 346, 350-351, 355, 357, 360-365, 373, 384-388, 393, 395, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 518, 532, 541, and 544-545 which are suppressed.

From each group of multiple events of transgenic plants with a specific recombinant DNA from Table 1 the event that produces the greatest enhancement in yield, water use efficiency, nitrogen use efficiency, enhanced cold tolerance, enhanced seed protein and enhanced seed oil is identified and progeny seed is selected for commercial development.

Progeny transgenic plants are selected from a population of transgenic cotton events under specified growing conditions and are compared with control cotton plants. Control cotton plants are substantially the same cotton genotype but without the recombinant DNA, for example, either a parental cotton plant of the same genotype that was not transformed with the identical recombinant DNA or a negative isoline of the transformed plant. Additionally, a commercial cotton cultivar adapted to the geographical region and cultivation conditions, i.e. cotton variety ST474, cotton variety FM 958, and cotton variety Siam L-23, are used to compare the relative performance of the transgenic cotton plants containing the recombinant DNA.

Transgenic cotton plants with enhanced yield and water use efficiency are identified by growing under variable water conditions. Specific conditions for cotton include growing a first set of transgenic and control plants under “wet” conditions, i.e. irrigated in the range of 85 to 100 percent of evapotranspiration to provide leaf water potential of −14 to −18 bars, and growing a second set of transgenic and control plants under “dry” conditions, i.e. irrigated in the range of 40 to 60 percent of evapotranspiration to provide a leaf water potential of −21 to −25 bars. Pest control, such as weed and insect control is applied equally to both wet and dry treatments as needed. Data gathered during the trial includes weather records throughout the growing season including detailed records of rainfall; soil characterization information; any herbicide or insecticide applications; any gross agronomic differences observed such as leaf morphology, branching habit, leaf color, time to flowering, and fruiting pattern; plant height at various points during the trial; stand density; node and fruit number including node above white flower and node above crack boll measurements; and visual wilt scoring. Cotton boll samples are taken and analyzed for lint fraction and fiber quality. The cotton is harvested at the normal harvest timeframe for the trial area. Enhanced water use efficiency is indicated by increased yield, improved relative water content, enhanced leaf water potential, increased biomass, enhanced leaf extension rates, and improved fiber parameters.

Example 5 Canola Transformation

This example illustrates plant transformation useful in producing the transgenic canola plants of this invention and the production and identification of transgenic seed for transgenic canola having enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil.

Tissues from in vitro grown canola seedlings are prepared and inoculated with overnight-grown Agrobacterium cells containing plasmid DNA with the gene of interest cassette and a plant selectable marker cassette. Following co-cultivation with Agrobacterium, the infected tissues are allowed to grow on selection to promote growth of transgenic shoots, followed by growth of roots from the transgenic shoots. The selected plantlets are then transferred to the greenhouse and potted in soil. Molecular characterizations are performed to confirm the presence of the gene of interest, and its expression in transgenic plants and progenies. Progeny transgenic plants are selected from a population of transgenic canola events under specified growing conditions and are compared with control canola plants. Control canola plants are substantially the same canola genotype but without the recombinant DNA, for example, either a parental canola plant of the same genotype that is not transformed with the identical recombinant DNA or a negative isoline of the transformed plant.

Transgenic canola plant cells are transformed with each of the recombinant DNA identified in Table 1. The above process is repeated to produce multiple events of transgenic canola plant cells that are transformed with recombinant DNA from each of the genes identified in Table 1. Events are designed to produce in the transgenic cells one of the proteins identified in Table 1, except the canola homologs of the proteins of SEQ ID NOs: 309, 311, 316, 324, 344, 346, 350-351, 355, 357, 360-365, 373, 384-388, 393, 395, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 518, 532, 541, and 544-545 which are suppressed. Progeny transgenic plants and seed of the transformed plant cells are screened for enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced seed protein and enhanced seed oil. From each group of multiple events of transgenic plants with a specific recombinant DNA from Table 1 the event that produces the greatest enhancement in yield, water use efficiency, nitrogen use efficiency, enhanced cold tolerance, enhanced seed protein and enhanced seed oil is identified and progeny seed is selected for commercial development.

Example 6 Homolog Identification

This example illustrates the identification of homologs of proteins encoded by the DNA identified in Table 1 which is used to provide transgenic seed and plants having enhanced agronomic traits. From the sequence of the homologs, homologous DNA sequence can be identified for preparing additional transgenic seeds and plants of this invention with enhanced agronomic traits.

An “All Protein Database” was constructed of known protein sequences using a proprietary sequence database and the National Center for Biotechnology Information (NCBI) non-redundant amino acid database (nr.aa). For each organism from which a polynucleotide sequence provided herein was obtained, an “Organism Protein Database” was constructed of known protein sequences of the organism; it is a subset of the All Protein Database based on the NCBI taxonomy ID for the organism.

The All Protein Database was queried using amino acid sequences provided herein as SEQ ID NO: 308 through SEQ ID NO: 614 using NCBI “blastp” program with E-value cutoff of 1e-8. Up to 1000 top hits were kept, and separated by organism names. For each organism other than that of the query sequence, a list was kept for hits from the query organism itself with a more significant E-value than the best hit of the organism. The list contains likely duplicated genes of the polynucleotides provided herein, and is referred to as the Core List. Another list was kept for all the hits from each organism, sorted by E-value, and referred to as the Hit List.

The Organism Protein Database was queried using polypeptide sequences provided herein as SEQ ID NO: 308 through SEQ ID NO: 614 using NCBI “blastp” program with E-value cutoff of 1e-4. Up to 1000 top hits were kept. A BLAST searchable database was constructed based on these hits, and is referred to as “SubDB”. SubDB is queried with each sequence in the Hit List using NCBI “blastp” program with E-value cutoff of 1e-8. The hit with the best E-value was compared with the Core List from the corresponding organism. The hit is deemed a likely ortholog if it belongs to the Core List, otherwise it is deemed not a likely ortholog and there is no further search of sequences in the Hit List for the same organism. For proteins of table 1 having an identified Pfam domain module, homology to reported homologs was further confirmed by searching the respective identified homologs for conservation of the Pfam domain module identified in the protein from table 1. Homologs from a large number of distinct organisms were identified and are reported below in table 7 with the SEQ ID NO of the DNA sequence corresponding to original protein query sequence and the identified homologs as [SEQ ID NO]: [Homolog SEQ ID N0s].

TABLE 7 Protein Sequences and their Homologs 1: 636 819 1097 1350 1889 1995 2165 2787 2937 3028 3574 3736 3793 3928 4331 4566 4851 5066 5207 5357 5398 5663 5664 6013 6476 6570 6701 6787 6959 7007 7057 7258 7401 7447 8148 8257 8679 8816 8900 8924 9039 9090 9392 9851 10181 10329 10383 10613 10783 10976 11004 11066 11131 11286 11480 11647 11722 11765 11832 12171 12412 12447 12798 12834 12898 12940 13177 13451 13570 13719 13950 14299 14438 14548 14592 14935 15659 15902 16223 16224 16225 16226 16227 16228 16274 16317 16550 16814 16848 17067 17087 17201 17262 17352 17689 18175 18176 18185 18519 18676 18855 18874 19060 19276 19656 19751 19922 20349 20723 21061 21081 21283 21397 21735 21976 22091 22248 22532 22808 23081 23178 23387 23483 23707 23922 23947 24001 24337 24585 24850 25090 25237 25290 25466 25663 25684 25788 25908 26445 26583 26823 27032 27073 27318 27324 27326 28043 28105 28199 28277 28306 28415 28508 28742 28774 29005 29247 29511 29652 30073 30230 30408 30456 30761 30816 30999 31063 31352 31385 31512 31559 31695 31898 32035 32178 32232 32337 32385 32394 32445 32691 32878 33131 33247 33249 33364 33479 34197 34394 34855 35042 35135 35209 35494 35573 35615 35629 35765 35871 35940 36015 36023 36173 2: 898 1260 1265 1375 1421 1533 1551 2768 3382 3404 3669 4434 5153 5179 5688 7146 7374 7885 8499 9364 11279 13414 13415 13416 13417 13418 13419 13420 13421 13422 16455 16779 16780 16781 16782 16783 18179 18180 18532 19184 19230 19766 20402 20438 21437 22781 23681 27774 27990 27999 28285 33033 34259 35255 35282 35285 35288 35289 36420 3: 456 457 470 712 881 1142 1361 2336 2631 2853 2989 3462 3491 3834 3927 4028 4048 4060 4127 4140 4160 4191 4241 4250 4302 5192 5679 5712 5811 5914 7872 7874 9042 9469 13359 13901 13913 16134 16173 17389 17790 17866 18577 18938 19220 19851 20421 20992 22719 23039 23691 24702 27436 27600 29693 31156 33797 36004 4: 344 385 4194 5: 741 1095 1126 1282 1869 2005 2063 2201 2860 3039 3267 3269 3340 3524 4426 6056 7596 8593 9054 9182 9183 9360 9694 9714 10214 11228 11363 11572 12507 13329 13330 13331 13332 13333 13334 13335 13347 13348 13349 13350 13351 13352 13425 13973 14651 15098 15291 15709 17686 17750 18147 18555 18750 18786 18813 19370 19457 19479 19838 19839 20150 20152 20184 20693 21241 21595 21596 22542 22643 22644 22645 22646 22798 23714 25078 25487 26417 26441 26498 26499 26500 27004 27025 28214 29144 30518 30709 30758 31167 31344 31394 31692 31734 32109 32138 32578 32959 33935 36167 36426 6: 775 1376 1383 1619 1972 2004 2037 2825 2866 4396 6439 9718 11312 11422 12356 12359 12365 12381 12402 12407 12443 12450 12455 12467 12468 12470 12472 12495 12498 12500 12501 16180 17480 17889 17895 18236 18975 19053 19298 19392 19761 19783 20005 20053 21580 21704 22589 22648 30963 30964 31241 31242 31243 31244 31245 31246 31247 31248 31249 31250 31251 33709 36349 7: 653 714 763 991 1289 2076 2340 2710 2905 3406 7933 11367 13848 15284 17753 19310 19456 19919 22778 31068 36330 8: 648 691 832 889 950 1035 1049 1085 1555 1747 1952 2074 2235 3471 4715 7929 9413 10227 10570 11271 13763 13766 13911 15477 17962 18934 18990 18991 20092 21463 22102 22819 25479 26945 28548 32656 33989 36086 36325 9: 1767 2508 2758 2873 2995 3173 3272 3961 5849 13687 13715 17896 18662 22881 10: 12216 13118 14734 14862 15330 17937 17977 18728 22144 22171 25713 30281 31805 33005 11: 733 1167 1803 1947 2134 2187 2226 2258 2325 2473 2532 3031 3165 3758 3934 3971 4005 4310 4453 4496 4523 4677 4710 4788 4797 4806 4827 4846 4847 4902 4910 4916 4942 4963 4971 5030 5063 5175 5235 5259 5269 5307 5446 5448 5512 5525 5543 5576 5639 5651 5654 5695 5848 5957 5967 5972 5994 6049 6098 6115 6126 6145 6172 6203 6209 6210 6211 6231 6234 6270 6287 6288 6289 6294 6295 6296 6301 6314 6330 6343 6377 6602 6607 6631 6661 6671 6696 6725 7028 7208 7233 7283 7428 7627 7655 7691 7695 7721 7899 8093 8095 8098 8121 8254 8278 8289 8307 8326 8352 8384 8523 8649 8664 8684 8737 8741 8750 8775 8778 8799 8800 8831 8835 8907 8946 8953 9003 9004 9013 9153 9161 9288 9340 9345 9518 9553 9559 9574 9678 9702 9719 9766 9792 9919 9922 10063 10084 10101 10113 10209 10298 10304 10312 10349 10440 10442 10486 10497 10509 10628 10646 10654 10721 10754 10773 10776 10840 10857 10861 10876 10882 10899 11004 11025 11041 11051 11072 11096 11110 11194 11255 11258 11450 11504 11527 11553 11597 11599 11603 11685 11734 11737 11825 11938 12012 12036 12056 12069 12073 12077 12080 12112 12136 12156 12161 12171 12187 12233 12250 12269 12291 12318 12405 12521 12522 12541 12561 12576 12615 12623 12649 12666 12683 12698 12728 12740 12850 12921 13038 13108 13184 13442 13465 13511 13546 13620 13718 13743 13786 13815 13860 13863 13868 13921 13981 13992 14021 14160 14167 14171 14180 14194 14255 14470 14632 14681 14688 14730 14749 14809 14819 14846 14882 14925 14933 14943 14976 14977 14983 15012 15031 15089 15090 15129 15139 15144 15168 15199 15327 15329 15335 15511 15527 15537 15698 15758 15772 15886 15910 15978 16079 16149 16158 16210 16267 16359 16363 16368 16371 16519 16557 16571 16633 16656 16674 16712 16720 16734 16736 16752 16875 17052 17064 17079 17105 17141 17146 17150 17186 17203 17225 17247 17402 17443 17452 17453 17454 17455 17456 17479 17508 17512 17536 17587 17592 17715 17894 17908 17922 17958 17997 18095 18102 18116 18126 18153 18258 18336 18338 18367 18383 18412 18421 18429 18452 18455 18469 18493 18502 18661 18676 18736 18762 18777 18784 18913 19019 19093 19105 19153 19465 19619 19699 19700 19733 19741 19894 20086 20089 20168 20172 20189 20213 20230 20256 20295 20307 20308 20379 20394 20556 20558 20590 20591 20611 20637 20638 20729 20741 20750 20752 20764 20772 20869 20874 20888 20962 20978 21024 21073 21252 21287 21374 21426 21623 21678 21684 21763 21779 21818 21852 22027 22032 22034 22094 22125 22164 22205 22224 22253 22301 22307 22320 22337 22359 22378 22389 22420 22435 22450 22486 22519 22627 22652 22818 22890 22891 22895 22966 23069 23075 23102 23161 23176 23253 23298 23356 23408 23424 23442 23541 23542 23547 23554 23592 23619 23736 23779 23780 23811 23815 23824 23833 23838 23851 23879 23907 23934 23942 23958 24097 24101 24106 24125 24137 24145 24369 24376 24450 24479 24495 24558 24612 24638 24651 24698 24850 24899 25020 25090 25149 25154 25189 25200 25208 25218 25237 25258 25265 25271 25290 25303 25309 25337 25358 25361 25407 25584 25612 25629 25647 25648 25732 25741 26255 26339 26407 26445 26552 26694 26717 26723 26858 26880 26891 27003 27069 27124 27213 27324 27331 27374 27377 27379 27409 27493 27526 27546 27558 27567 27606 27626 27649 27650 27660 27695 27698 27707 27802 27866 27889 27912 27959 28049 28068 28113 28132 28174 28269 28377 28443 28478 28679 28719 28801 28856 28866 28881 28906 28914 29014 29022 29033 29106 29139 29160 29168 29242 29266 29285 29316 29337 29339 29404 29408 29454 29538 29566 29575 29598 29713 29716 29735 29739 29767 29773 29789 29793 29809 29826 29933 29935 29949 29968 30016 30057 30102 30125 30141 30162 30173 30188 30226 30230 30234 30277 30309 30368 30408 30429 30456 30472 30489 30540 30551 30561 30563 30609 30638 30639 30654 30683 30792 30793 30866 30878 30884 30904 30924 30967 31063 31134 31215 31216 31219 31375 31482 31490 31602 31733 31810 32008 32013 32023 32049 32051 32055 32234 32346 32358 32381 32398 32422 32462 32494 32521 32529 32653 32664 32669 32722 32753 32762 32772 32776 32789 32798 32855 32875 32889 32986 33002 33059 33119 33146 33152 33197 33219 33286 33300 33316 33329 33351 33442 33460 33486 33526 33621 33643 33663 33690 33697 33699 33772 33845 33846 33847 33848 33849 33850 33929 33930 33947 34024 34031 34038 34063 34085 34161 34164 34199 34216 34238 34251 34286 34288 34317 34334 34348 34399 34545 34598 34688 34697 34778 34808 34832 34879 34895 34900 34936 34972 34996 35057 35083 35111 35181 35229 35263 35268 35362 35377 35428 35470 35671 35679 35704 35708 35720 35733 35738 35770 35806 35842 35865 35892 35946 35949 36060 36083 36102 36124 36180 36206 36250 36267 36275 36398 12: 3551 4778 5210 5604 7474 8618 9196 10382 11155 11645 12560 12587 12810 12835 13066 13172 13568 13630 14570 15055 15890 16081 16563 17858 18083 18323 18839 20636 21261 21978 22393 23236 23580 24043 24809 25035 25102 25138 25306 25786 26437 27283 28010 28108 28725 29361 29646 29878 30100 30657 31040 31866 32676 34201 35216 35792 13: 4912 5245 5316 5329 6070 6515 6660 7632 7661 7690 7728 7864 8199 8295 8840 8957 8999 9165 9520 10348 10398 10414 10433 10463 10996 11052 11091 11132 11174 11457 11488 11625 11824 12140 12165 12225 12277 12548 12565 12715 12851 12866 12922 12930 12937 13095 13545 13595 13619 13647 13657 13778 13983 14473 14643 14733 14788 14801 15116 15379 15490 15541 15697 16023 16275 16387 16414 16443 16659 17092 17113 17899 17965 18161 18274 18275 18398 18438 18731 18765 19738 19755 20260 20850 20912 21251 21770 21778 22001 22022 22077 22099 22172 22267 22368 22461 23199 23240 23252 23609 23765 23993 24126 24132 24152 24664 25024 25343 25362 25404 25715 25803 25839 25961 26610 26670 27096 27557 27617 27641 27796 28079 28087 28101 28802 28841 28848 28965 29889 29907 30283 30294 30329 30335 30546 31591 31792 31986 32520 32522 32536 32619 32737 32965 33281 33532 33778 33832 34013 34236 34673 34791 34998 35123 35245 35604 35728 14: 451 452 467 2486 2567 4431 4518 4563 4590 4660 4684 4686 4727 4786 4809 4862 4864 5005 5019 5036 5055 5141 5201 5266 5284 5313 5324 5355 5371 5387 5479 5538 5551 5573 5609 5908 6074 6079 6132 6156 6173 6204 6233 6341 6360 6361 6497 6558 6568 6592 6620 6689 6733 6742 6747 6806 6874 6875 6916 7019 7069 7123 7128 7157 7177 7239 7457 7473 7636 7664 7680 7684 7731 8166 8224 8235 8299 8419 8506 8609 8637 8730 8771 8797 8869 9241 9325 9524 9529 9610 9759 9814 9830 9849 9912 9931 9972 9992 9999 10052 10334 10363 10467 10667 10677 10700 10738 10772 10782 10813 10850 10885 10894 10975 11031 11154 11460 11528 11569 11636 11704 11752 11766 11886 11930 11934 12016 12050 12201 12210 12211 12222 12323 12331 12352 12471 12530 12555 12586 12639 12691 12759 12763 12843 12916 13061 13076 13083 13145 13176 13195 13211 13429 13557 13625 13759 13967 14169 14281 14304 14359 14360 14402 14481 14512 14564 14634 14709 14715 14726 14826 14855 14864 14957 14979 15057 15207 15228 15342 15370 15517 15914 15936 15969 16069 16293 16441 16575 16593 16692 16718 16823 16851 16861 17036 17102 17228 17337 17351 17418 17421 17516 17582 17583 17727 17728 17912 17931 17966 17987 17988 18082 18106 18125 18309 18358 18363 18391 18476 18484 18707 18732 18843 18885 19620 19666 19703 19759 20284 20560 20573 20596 20602 20626 20696 20883 20911 20966 21004 21160 21210 21671 21753 21802 21987 22121 22145 22177 22180 22238 22278 22303 22326 22332 22341 22382 22416 22463 22504 22526 23086 23138 23263 23434 23481 23488 23520 23536 23588 23631 23760 24003 24041 24091 24304 24323 24392 24433 24490 24523 24549 24559 24647 24717 24761 24796 24797 24819 24827 24828 24829 24879 24915 25003 25007 25029 25033 25176 25231 25346 25368 25372 25396 25398 25492 25520 25747 25775 25827 25907 26249 26301 26359 26396 26571 26582 26738 26779 26785 26830 27074 27273 27355 27368 27369 27382 27398 27471 27495 27553 27621 27676 27751 27769 27772 27963 27968 28061 28092 28483 28502 28538 28555 28601 28602 28623 28696 28738 28806 28882 28901 28947 28994 29118 29119 29177 29272 29298 29401 29431 29456 29464 29547 29548 29582 29609 29637 29645 29660 29745 29813 29835 29883 29963 29980 29989 30054 30068 30155 30195 30256 30292 30332 30643 30876 30991 31019 31049 31161 31530 31809 31812 31840 31841 31842 31843 32072 32174 32238 32289 32377 32433 32443 32458 32501 32540 32564 32565 32567 32568 32621 32631 32639 32646 32673 32706 32872 32967 32999 33082 33185 33195 33293 33332 33381 33417 33456 33537 33540 33572 33635 33782 33833 34086 34104 34109 34131 34169 34247 34258 34287 34304 34308 34322 34423 34476 34512 34544 34615 34677 34685 34714 34716 34765 34781 34794 34796 34813 34826 34881 34942 34953 34979 35002 35046 35100 35103 35113 35121 35136 35142 35182 35395 35400 35427 35489 35496 35525 35526 35527 35546 35552 35572 35694 35829 35849 35955 35978 36111 36129 36187 36224 36289 36367 36383 15: 323 793 812 880 904 916 1016 1377 1453 1492 1516 1626 1764 1790 1791 1949 1950 2042 2154 2458 2616 2703 2819 2877 2911 2914 2948 3015 3134 3154 3260 3261 3270 3297 3303 3329 3365 3451 3852 3855 3915 4116 4129 4142 4167 4207 4246 4279 4370 4393 4460 4471 4500 4505 4547 4583 4595 4611 4612 4613 4616 4617 4663 4670 4673 4675 4811 4824 4858 4955 4983 5001 5003 5006 5009 5015 5018 5020 5025 5028 5029 5031 5032 5044 5046 5057 5071 5078 5108 5232 5335 5364 5382 5406 5441 5465 5477 5558 5585 5589 5599 5632 5676 5840 5906 5939 5948 5952 5961 5982 6164 6167 6176 6207 6363 6365 6537 6538 6555 6579 6608 6676 6680 6690 6692 6697 6703 6714 6722 6724 6755 6758 6770 6775 6779 6819 6913 6970 6979 7021 7034 7043 7045 7060 7114 7117 7135 7166 7173 7183 7206 7222 7225 7261 7266 7272 7305 7405 7406 7407 7413 7429 7432 7455 7456 7586 7853 7854 7863 8128 8139 8161 8169 8181 8182 8185 8186 8226 8229 8230 8252 8261 8268 8319 8348 8418 8492 8513 8542 8608 8610 8628 8631 8658 8677 8680 8696 8756 8764 8828 8844 8848 8849 8850 8899 8901 8974 9118 9121 9140 9145 9179 9192 9226 9227 9234 9235 9258 9465 9575 9595 9614 9615 9623 9624 9751 9769 9772 9778 9780 9805 9808 9813 9835 9920 9924 9991 10020 10056 10115 10154 10362 10364 10366 10371 10396 10444 10449 10450 10489 10505 10510 10515 10552 10553 10566 10599 10610 10617 10621 10632 10678 10733 10799 10824 10841 10853 10878 10880 10901 10905 10986 11026 11092 11106 11129 11203 11231 11385 11531 11537 11567 11694 11700 11705 11709 11715 11747 11784 11794 11801 11806 11810 11812 11816 11817 11828 11923 11942 11947 11948 11966 11997 12019 12081 12125 12213 12263 12271 12280 12303 12305 12404 12559 12625 12641 12657 12663 12725 12732 12733 12750 12774 12809 12885 12924 12947 13019 13036 13051 13088 13126 13130 13147 13214 13215 13258 13259 13343 13455 13457 13461 13474 13506 13509 13581 13640 13692 13748 13946 14012 14195 14253 14256 14264 14267 14286 14392 14430 14589 14787 14800 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27744 27886 27905 27973 28318 28490 28495 28615 28628 28713 28724 28804 28982 29161 29178 29230 29239 29257 29425 29494 29497 29608 29811 29888 29925 30011 30014 30019 30053 30056 30069 30085 30215 30236 30445 30620 30640 31013 31039 31124 31154 31232 31513 31780 31849 31918 32036 32060 32065 32107 32187 32237 32261 32263 32288 32300 32343 32348 32360 32461 32481 32493 32505 32535 32647 32658 32666 32667 32784 32884 32939 32975 32983 33132 33155 33160 33164 33198 33483 33489 33499 33533 33779 34070 34089 34158 34162 34187 34255 34314 34337 34382 34397 34405 34446 34483 34490 34532 34553 34628 34672 34696 34728 34747 34779 34969 34997 35198 35205 35217 35275 35517 35636 35706 35735 35768 35769 35861 35880 35897 35963 36088 36122 36123 39: 426 946 40: 761 821 978 1165 2748 2749 2816 3174 3561 3822 4144 4247 7326 8364 8365 8366 8861 8862 9122 9733 11408 13752 13758 13862 13953 13963 14127 16775 17809 17859 18634 18748 18785 18799 19284 19401 20045 20444 21091 21631 21827 22579 23219 23713 24157 24158 25649 25909 27451 28836 32951 34582 34857 41: 309 411 898 905 915 1533 1595 1792 2768 3382 3404 3669 4434 5153 5688 6982 7146 7374 7885 9364 10152 10153 11279 15067 16455 16779 16780 16781 16782 16783 18532 19184 19230 19766 20402 21437 22781 24696 25892 26049 27990 33033 34259 35288 35289 36420 42: 309 348 410 411 412 630 898 905 915 1533 1595 1792 2768 3382 3404 3669 4434 5153 5688 6982 7146 7374 7885 9364 10152 10153 11279 15067 16455 16779 16780 16781 16782 16783 18532 19184 19230 19766 20402 21437 22781 24696 25892 26049 27990 33033 34259 35288 35289 36420 43: 762 1070 1190 1272 1505 1924 2220 4883 5276 5689 6016 6047 6906 6947 6976 9448 10058 10194 11275 13784 13851 14391 14392 15431 15638 15639 15640 15641 15642 15643 15644 15645 15646 15647 15648 15685 15686 16312 17056 17771 18566 19255 19343 19527 19971 20097 21418 22878 24160 24742 25052 25867 26091 26288 26489 26595 27761 28272 28299 28387 30624 30753 31744 31903 32119 32941 33063 33064 34566 36008 36422 44: 1671 2465 3294 5450 5774 6626 7964 8053 10369 10714 11634 12842 12915 13048 13556 13626 13764 13780 13897 15202 15975 19324 19790 21532 22559 23091 23262 23566 24522 24842 25250 25589 26597 28971 29367 29373 29612 29882 31853 32615 32701 35206 35411 45: 977 1038 1039 1148 1382 1602 1639 1690 1709 1732 1793 1844 1856 1946 2035 2047 2144 2145 2151 2152 2199 2209 2261 2280 2283 2300 2316 2318 2341 2361 2364 2383 2389 2405 2435 2469 2530 2602 2648 2659 2664 2670 2705 2731 2735 2776 2780 2786 2933 2986 3020 3024 3025 3057 3103 3132 3150 3157 3179 3181 3201 3280 3309 3313 3341 3485 3511 3519 3529 3571 3673 3804 3805 3833 3932 3992 3993 4032 4096 4221 4483 5285 5286 5287 5290 5685 5716 5717 5718 5719 5751 5805 5806 5823 5824 5825 5826 5855 5864 6405 6410 6424 6437 6483 6646 7882 7896 7902 7924 7925 7926 7988 7989 7999 8000 8059 8389 8390 8391 8912 9441 9724 9898 10159 10236 10268 11263 11398 11399 11407 11901 12024 12025 12997 13104 13776 14238 14994 14995 14996 15077 15298 15299 15493 15505 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31142 31178 31277 31411 31436 31578 31579 31580 31581 31871 32593 32594 32913 32914 32915 33034 34921 35302 35535 36228 36239 36421 46: 977 1038 1039 1148 1382 1602 1639 1690 1709 1732 1793 1844 1856 1946 2035 2047 2144 2145 2151 2152 2199 2209 2261 2280 2283 2300 2316 2318 2341 2361 2364 2383 2389 2405 2435 2469 2530 2602 2648 2659 2664 2670 2705 2731 2735 2776 2780 2786 2933 2986 3020 3024 3025 3057 3103 3132 3150 3157 3179 3181 3201 3280 3309 3313 3341 3485 3511 3519 3529 3571 3673 3804 3805 3833 3932 3992 3993 4032 4096 4221 4483 5285 5286 5287 5290 5685 5716 5717 5718 5719 5751 5805 5806 5823 5824 5825 5826 5855 5864 6405 6410 6424 6437 6483 6646 7882 7896 7902 7924 7925 7926 7988 7989 7999 8000 8059 8389 8390 8391 8912 9441 9724 9898 10159 10236 10268 11263 11398 11399 11407 11901 12024 12025 12997 13104 13776 14238 14994 14995 14996 15077 15298 15299 15493 15505 15520 15737 15738 15739 15740 15741 16001 16045 16168 16187 16190 16457 16507 16614 16701 17136 17482 17762 17820 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36228 36239 36421 47: 847 874 1056 1405 1721 1857 1873 1977 1978 2077 2496 2730 3014 3244 3458 4438 4499 4520 4879 5815 6048 6366 7764 7765 7766 7767 7768 7769 7770 8572 9077 9113 9419 9452 10167 10218 11251 11252 11346 11347 12429 12960 14034 14174 14176 14237 15631 15985 17874 18646 18704 18819 19090 19256 19527 19897 19898 20574 21541 22772 23157 23568 24160 25478 25686 26275 26282 26309 26435 26501 26989 27104 27754 28138 28190 28258 28454 30401 30743 31060 31224 31342 31935 32095 32471 34066 34595 34820 48: 1195 1211 1244 1485 1528 1574 1684 2268 2574 2963 2975 3097 3127 3468 5113 5694 6534 8877 9135 9190 11352 13667 16174 16496 17477 17600 17776 18568 18696 19114 19190 20373 21425 31775 35992 35993 35994 35995 35996 36227 49: 782 1561 1934 1936 2192 2257 2271 2393 2446 2700 2953 3957 3959 4405 5080 5110 5703 5856 5866 5977 6006 6019 6465 6923 6994 7054 7506 8794 9466 10141 10216 10662 10804 11365 11500 12525 12736 13025 13251 13804 13805 13919 14038 14244 14410 14611 14763 14797 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17000 17043 17131 17168 17231 17378 17567 17598 17724 17830 17948 18005 18074 18112 18157 18225 18288 18375 18407 18488 18510 18603 18652 18670 18735 18766 18794 18837 18871 19238 19611 19713 19852 20191 20219 20250 20291 20633 20768 20830 20881 20944 20977 20991 21033 21122 21237 21266 21277 21288 21344 21380 21676 21751 21760 21810 22008 22075 22106 22176 22209 22297 22353 22376 22443 22493 22515 22571 22687 23109 23148 23314 23349 23443 23456 23511 23628 23703 23735 23787 23841 23876 23902 23923 23931 23991 24112 24292 24383 24407 24488 24564 24646 24769 24830 24868 24884 24956 24983 25012 25058 25107 25141 25329 25373 25416 25477 25497 25564 25587 25615 25624 25669 25750 25807 25841 25899 25960 26089 26196 26261 26311 26372 26399 26414 26565 26688 26711 26766 26788 26824 26887 27008 27113 27274 27295 27362 27408 27462 27503 27684 27705 27851 27875 27941 27966 28048 28077 28166 28211 28293 28366 28414 28484 28507 28533 28646 28695 28765 28789 28874 28964 28998 29083 29174 29274 29346 29411 29439 29461 29567 29599 29626 29840 29913 29919 29960 29987 30034 30099 30183 30239 30265 30303 30344 30397 30418 30425 30451 30478 30531 30565 30615 30630 30714 30796 30856 30888 30911 30928 30933 30977 31056 31109 31203 31373 31423 31479 31685 31823 31956 32058 32135 32195 32233 32287 32387 32399 32469 32514 32554 32708 32728 32783 32882 32980 33041 33154 33213 33248 33282 33334 33453 33521 33553 33609 33672 33766 33864 34015 34064 34195 34242 34284 34345 34410 34471 34547 34603 34684 34689 34766 34816 34885 34905 34948 34978 35035 35093 35162 35213 35279 35387 35450 35490 35593 35630 35666 35780 35899 35914 36059 36131 36216 36240 36354 36389 291: 2040 2204 2456 3374 3595 4026 4037 4408 6181 6429 7957 9460 11249 11310 13934 14072 17793 19405 19817 20074 21459 21460 21524 21570 21571 22568 22705 27438 27439 292: 293: 639 1116 1436 1537 2691 3175 3465 3902 4131 5710 7349 7788 8385 10538 11216 11218 11370 15749 18640 18648 18933 19583 19979 21477 23918 25817 30514 33507 294: 1013 1521 2093 2191 2216 2217 3754 4332 5778 6059 6925 7972 8563 8819 8952 9014 9089 9152 12680 13210 13990 14403 15530 15687 16344 16516 16517 18861 18995 19879 21617 22020 22728 26017 26958 27034 27242 28134 28197 28238 28361 28372 28461 29015 30545 30697 31362 31438 31896 32131 32916 32964 295: 2370 2478 2521 2711 2868 2998 3104 3241 3873 3903 4010 4089 4113 4497 4629 4655 4801 5052 5358 5401 5457 5555 5607 5784 6061 6147 6394 6403 6415 6544 6573 6623 6684 6809 6905 7010 7085 7160 7262 7452 8004 8005 8071 8192 8280 8515 8590 8666 8748 9248 9552 9566 9747 9829 10021 10086 10295 10378 10488 10627 10828 10866 10972 11542 11595 11641 11696 11760 11778 12014 12544 12722 12841 12976 13136 13447 13562 14004 14248 14428 14445 14808 14927 15028 15262 15324 15882 16063 16076 16142 16333 16570 16821 17049 17178 17239 17353 17394 17442 17494 17846 17847 18314 18473 18655 18891 19648 20345 20614 20763 20909 21027 21064 21137 21257 21565 21566 21605 21697 21731 21870 21932 21981 22263 22329 22342 22394 23123 23158 23265 23379 23474 23525 23774 23943 24011 24073 24372 24693 24786 25147 25215 25289 25508 25782 25851 25943 26210 26376 26542 27026 27380 27410 28013 28093 28189 28248 28332 28480 29264 29324 29449 29555 29588 29649 29723 29880 30015 30076 30150 30347 30703 30867 30996 31229 31388 31491 31960 31997 32101 32127 32199 32266 32379 32454 32699 33096 33205 33343 33377 33462 33555 34055 34163 34213 34272 34323 34430 34475 34546 34632 34707 34965 35041 35073 35167 35185 35389 35495 35580 35890 35929 35936 36097 36213 36415 296: 790 939 1402 1446 1833 2210 2433 2596 2704 2809 2851 3086 3250 3358 3389 3411 3457 3634 3697 3894 4227 5812 7928 7958 8046 8061 13711 13977 18169 20413 21462 21526 21662 22703 22843 22990 23302 23807 24697 26813 32576 34746 297: 917 920 1102 1103 1171 1212 1299 1406 1657 1713 1780 1908 1941 2114 2238 2244 2353 2412 2588 2706 2779 2827 2943 3005 3056 3082 3239 3326 3370 3378 3438 3486 3591 3623 3652 3674 3704 3712 3815 3825 3874 3958 3969 4024 4094 4107 4240 4259 4270 4297 4309 4391 5692 5830 6409 6853 7889 7948 8003 9432 12370 13883 14016 14032 14069 14096 16126 16135 16151 16154 17388 17497 17653 17754 18557 18567 18925 18936 19025 19156 19395 19603 19813 19835 20071 20384 20389 20406 21326 21350 21588 21610 22570 22838 22873 22930 22956 22987 29384 29854 31872 298: 530 606 623 742 1622 2031 2416 2550 2657 2755 3038 3072 3117 3163 3195 3228 3554 3682 3741 3783 3884 3885 4117 4163 4196 4345 4392 4448 4465 4705 4716 4717 4742 4748 4754 5120 5187 5189 5852 6389 6404 6432 6493 6983 8484 8485 8486 8487 8498 9120 9181 9648 9716 10532 10533 10592 11219 11229 11420 12705 12706 12707 12987 12988 13322 13323 13947 14025 14655 15553 15793 15794 16008 16289 17780 18572 18792 18947 19027 19360 19489 20036 20046 20318 20450 21087 21223 21224 21225 21226 21227 21228 21340 21446 21915 21916 21917 22578 22606 23063 23690 24161 24536 24537 24538 24689 24690 24727 24757 24803 24922 24924 24934 25310 25653 25938 25939 26440 26504 26530 26667 26668 26901 26902 26903 26906 26907 26908 26913 26919 27823 28562 29847 30097 31122 31123 31157 31188 31192 31320 31432 31533 31598 31829 31830 31978 32600 32826 34923 35063 35251 36315 36316 36317 36318 299: 530 605 623 660 742 744 845 859 1025 1026 1599 1622 1687 1820 1925 1994 2031 2120 2416 2550 2657 2755 3038 3072 3117 3163 3195 3228 3554 3682 3741 3783 3884 3885 4117 4163 4196 4345 4392 4448 4465 4717 4740 4741 4742 4748 4889 5187 5189 5483 5852 6389 6404 6432 6493 6503 8484 8485 8486 8487 8498 9044 9120 9181 9653 9716 10532 10533 11219 11229 11420 11483 12705 12706 12707 12987 12988 13322 13323 14054 14087 14655 15510 15722 15793 15794 16280 16698 16708 17692 17780 18267 18270 18572 18792 18947 18961 19027 19067 19282 19360 19489 19730 19793 20036 20046 20318 20450 20528 20927 20929 20930 21087 21222 21223 21224 21225 21227 21307 21308 21309 21340 21446 21915 21916 21917 22469 22472 22578 22606 23011 23063 23676 23684 23690 24161 24321 24536 24537 24538 24541 24689 24690 24727 24757 24803 24919 24922 24924 24934 25310 25653 25938 25939 26064 26427 26428 26429 26440 26491 26504 26530 26901 26902 26905 26906 26907 26913 26919 27046 27586 27588 27589 27823 28280 28374 28562 29847 30097 30629 31071 31122 31123 31157 31188 31192 31278 31315 31316 31320 31418 31432 31533 31565 31598 31753 31754 31829 31830 31831 31978 31980 31981 31982 31983 32600 32826 32850 33822 33966 34574 34923 35063 35248 36301 36302 36308 36315 36316 36317 36318 300: 967 1454 2468 2900 3185 3630 4004 5773 6185 9053 10220 10221 10222 11068 13896 14902 17524 17843 21551 21713 23860 26528 26916 26917 26946 31184 31185 36334 301: 933 992 2437 2553 2614 2637 2745 3492 3566 3640 3660 3716 3867 3920 4092 4281 4371 4374 5731 5767 6190 6458 7960 10548 13935 14136 14902 17659 17801 18170 18611 18793 18807 20364 20454 21490 21536 21545 21713 26528 26915 26916 26917 27140 29377 29378 33803 36235 36334 302: 303: 2071 304: 908 938 1053 1243 1370 1472 2228 2992 3424 4099 6869 11234 13208 17834 19458 20078 20375 21312 22816 23030 23667 305: 615 635 662 665 690 863 1008 1021 1087 1185 1296 1369 1380 1399 1545 1597 1635 1664 1678 1681 1692 1771 1872 1898 1929 1945 1974 1991 2021 2051 2088 2089 2090 2110 2116 2122 2132 2224 2248 2298 2335 2455 2460 2506 2561 2564 2610 2683 2689 2694 2695 2765 2784 2811 2839 2840 2847 2849 2867 2894 2944 3013 3224 3377 3388 3446 3450 3484 3531 3537 3546 3547 3560 3577 3593 3659 3707 3843 3983 3984 4022 4054 4057 4133 4141 4232 4248 4293 4402 4403 4976 4977 4978 4980 5171 5655 5750 5862 5871 6498 7606 7908 7915 7916 7931 7946 7969 7970 7983 8030 8049 8817 8942 8943 9347 9361 9362 9380 9434 9646 9942 9943 9944 9945 9946 9947 9948 9949 10125 10126 10127 10137 10266 10279 10283 10881 11067 11069 11199 11200 11262 11330 11838 13222 13223 13555 14561 15281 15628 15629 15800 16068 16148 16152 16349 16613 17021 17395 17397 17533 17699 17757 17768 18264 18442 18543 18564 18598 18667 18668 18949 18965 18968 19029 19071 19072 19165 19167 19170 19203 19225 19243 19245 19288 19292 19341 19373 19431 19764 19849 19858 19881 20012 20016 20049 20369 20388 20430 20642 20644 20652 20984 20985 20986 20987 21093 21245 21332 21402 21412 21415 21416 21487 21516 21600 21615 21616 21801 22651 22792 22841 22889 22913 22949 22968 22974 22976 22980 23687 23913 24075 24940 26053 26442 26603 26605 27042 30139 32158 32925 32926 33230 33798 33945 34365 34369 34370 34371 34372 34373 34374 34375 34376 35477 306: 614 677 693 857 1139 1412 1536 1794 2007 2025 2447 2493 2699 2762 3840 4046 4253 4315 4458 4472 4498 4504 4565 4575 4576 4580 4581 4610 4625 4854 4888 4940 4948 4988 4989 4990 5002 5014 5017 5024 5047 5048 5072 5077 5083 5086 5092 5109 5115 5117 5366 5370 5407 5409 5559 5744 5950 5969 5971 6116 6177 6179 6342 6519 6580 6582 6686 6702 6708 6709 6726 6730 6739 6761 6762 6773 6793 6796 6803 6814 7008 7134 7136 7154 7178 7182 7192 7221 7243 7253 7254 7255 7267 7427 7500 7501 7502 7503 7587 7588 7741 7865 7944 8020 8042 8079 8134 8135 8136 8137 8138 8140 8147 8154 8155 8157 8158 8180 8183 8191 8225 8228 8241 8386 8406 8517 8612 8613 8626 8632 8661 8769 8875 8913 8948 8968 9219 9251 9540 9541 9571 9572 9682 9708 9855 9858 9902 9923 9971 9979 9993 10000 10018 10092 10120 10160 10161 10191 10390 10448 10455 10527 10612 10650 10652 10664 10673 10681 10812 10816 10825 10860 11308 11309 11316 11430 11498 11576 11654 11717 11719 11720 11721 11802 11822 11852 11885 12304 12355 12526 12618 12769 12828 12909 13016 13017 13020 13021 13022 13125 13158 13362 13431 13440 13462 13476 13497 13578 13637 13881 14156 14234 14252 14257 14268 14282 14288 14292 14458 14459 14464 14551 14700 14740 14803 14900 14901 14967 14971 15094 15131 15206 15220 15229 15260 15280 15350 15486 15717 15920 15921 15939 15941 16028 16109 16246 16257 16420 16421 16422 16423 16424 16425 16426 16427 16428 16530 16574 16582 16583 16608 16797 16837 16869 17023 17096 17097 17100 17101 17163 17207 17265 17370 17379 17407 17441 17462 17474 17704 17945 18027 18252 18253 18254 18255 18256 18257 18301 18453 18740 18741 19191 19496 19597 19626 19634 19646 19814 19815 20011 20173 20312 20315 20475 20477 20564 20586 20618 20620 20742 20767 20778 20803 20849 20990 21069 21140 21164 21165 21263 21727 21833 21873 21904 21921 21971 22059 22082 22086 22237 22264 22316 22377 22379 22543 22544 22713 22714 23006 23022 23155 23166 23168 23182 23272 23343 23354 23368 23401 23410 23439 23440 23460 23478 23502 23532 23546 23562 23605 23613 23662 23668 23719 23927 24006 24028 24033 24322 24331 24364 24386 24448 24610 24723 24740 24800 24807 24874 24882 24914 24973 24974 25086 25097 25122 25125 25126 25133 25169 25187 25214 25216 25251 25263 25266 25300 25301 25305 25326 25429 25470 25486 25515 25534 25664 25777 25794 25815 25823 25849 25862 25888 25889 25895 25896 25945 26070 26074 26077 26241 26310 26506 26544 26570 26576 26579 26581 26778 26794 26856 26935 26964 27091 27102 27202 27244 27605 27737 27827 27943 27950 28021 28110 28194 28226 28367 28368 28410 28473 28475 28631 28770 28771 28832 28833 28852 28934 28975 29271 29336 29402 29472 29474 29477 29485 29487 29495 29503 29533 29606 29618 29665 29667 29679 29872 29945 30052 30083 30147 30176 30325 30423 30579 30650 30682 30736 30829 30881 30949 31007 31025 31099 31155 31158 31254 31276 31370 31466 31467 31468 31502 31526 31560 31714 31749 31836 31837 31838 31839 31900 31962 31963 31964 32024 32031 32052 32063 32068 32223 32245 32248 32256 32349 32367 32378 32407 32419 32450 32474 32492 32614 32640 32684 32712 32782 32885 32896 32910 32911 32946 33051 33077 33109 33120 33148 33158 33163 33259 33260 33261 33262 33276 33278 33325 33368 33374 33376 33378 33392 33429 33457 33997 34115 34122 34183 34267 34447 34513 34579 34612 34731 34911 35022 35051 35090 35115 35171 35173 35196 35218 35228 35393 35521 35545 35558 35559 35562 35600 35755 35789 35790 35797 35809 35820 35825 35828 35854 35918 35961 35962 36089 36115 36134 36135 36148 36160 36166 36200 36273 36284 36360 36361 36369

Recombinant DNA constructs are prepared using the DNA encoding each of the identified homologs and the constructs are used to prepare multiple events of transgenic corn, soybean, canola and cotton plants as illustrated in Examples 2-5. Plants are regenerated from the transformed plant cells and used to produce progeny plants and seed that are screened for enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil. From each group of multiple events of transgenic plants with a specific recombinant DNA for a homolog the event that produces the greatest enhancement in yield, water use efficiency, nitrogen use efficiency, enhanced cold tolerance, enhanced seed protein and enhanced seed oil is identified and progeny seed is selected for commercial development.

Example 7 Consensus Sequence

This example illustrates the identification of consensus amino acid sequence for the proteins and homologs encoded by DNA that is used to prepare the transgenic seed and plants of this invention having enhanced agronomic traits.

ClustalW program was selected for multiple sequence alignments of the amino acid sequence of SEQ ID NO: 311, 314, 316, 333, 344, 346, 365, 373, 383-386, 404-405, 426, 449, 461, 466, 478, 488, 490-491, 512, 541, 543-544, 559, 579, 583, 603, and 610-611 and their homologs. Three major factors affecting the sequence alignments dramatically are (1) protein weight matrices; (2) gap open penalty; (3) gap extension penalty. Protein weight matrices available for ClustalW program include Blosum, Pam and Gonnet series. Those parameters with gap open penalty and gap extension penalty were extensively tested. On the basis of the test results, Blosum weight matrix, gap open penalty of 10 and gap extension penalty of 1 were chosen for multiple sequence alignment.

The consensus amino acid sequence can be used to identify DNA corresponding to the full scope of this invention that is useful in providing transgenic plants, for example corn and soybean plants with enhanced agronomic traits, for example improved nitrogen use efficiency, improved yield, improved water use efficiency and/or improved growth under cold stress, due to the expression in the plants of DNA suppressing a protein with amino acid sequence identical to the consensus amino acid sequence.

The SEQ ID NOs for the identified consensus sequences are reported in table 8 below and the full consensus sequences are provided in the attached sequence listing.

TABLE 8 PEP SEQ ID NO Consensus SEQ ID NO 311 36447 314 36448 316 36449 333 36450 344 36451 346 36452 365 36453 373 36454 383 36455 384 36456 385 36457 386 36458 404 36459 405 36460 426 36461 449 36462 461 36463 466 36464 478 36465 488 36466 490 36467 491 36468 512 36469 541 36470 543 36471 544 36472 559 36473 579 36474 583 36475 603 36476 610 36477 611 36478

Example 9 Identification of Amino Acid Domain by Pfam Analysis

This example illustrates the identification of domain and domain module by Pfam analysis.

The amino acid sequence of the expressed proteins that are shown to be associated with an enhanced trait were analyzed for Pfam protein family against the current Pfam collection of multiple sequence alignments and hidden Markov models using the HMMER software. The Pfam protein domains and modules for the proteins of SEQ ID NOs: 308-310, 312-313, 315, 317-332, 334-343, 345, 347-364, 366-372, 374, 382, 387-403, 406-412, 414-425, 427-448, 450-460, 462-465, 467-477, 479-482, 484-487, 493-511, 513-539, 542, 545-558, 560-578, 580-582, 584-598, 600-602, 604-608, and 612-614 are shown in Tables 9, 10 and 11. The Hidden Markov model databases for the identified patent families are also available from the Pfam consortium (ftp.sanger.ac.uk/pub/databases/Pfam/) allowing identification of other homologous proteins and their cognate encoding DNA to enable the full breadth of the invention for a person of ordinary skill in the art. Certain proteins are identified by a single Pfam domain and others by multiple Pfam domains. The function of the identified Pfam domains in proteins providing an enhanced trait in plants was verified by searching identified homologs for the conservation of the identified Pfam domains. The score value for the identified Pfam domains in sequences from table 1 and the minimum score value for the Pfam domain between a protein from table 1 and its identified homologs are reported below in table 9.

TABLE 9 Pfam annotation PEP SEQ ID Pfam domain Minumun NO name Begin Stop Score Score E-value 308 Aminotran_1_2 69 470 60.3 −40.6 7.20E−15 309 Homeobox 65 119 67.2 −2.9 6.20E−17 309 HALZ 120 164 45.5 25 2.10E−10 310 bZIP_1 108 171 40.6 27.4 6.30E−09 310 bZIP_2 108 162 42.2 18.1 2.10E−09 312 Peptidase_S10 39 504 699 −109.9 4.00E−207 313 SBP 103 181 167.3 41.4 4.60E−47 315 GDPD 436 728 154.9 −14.9 2.50E−43 317 DUF1292 111 180 19.6 16.5 4.10E−05 318 Aminotran_1_2 127 473 234 −30.7 3.70E−67 319 FAD_binding_4 50 191 14.8 −6.6 0.00012 319 Lact-deh-memb 279 570 657.8 75.1 9.70E−195 320 Fe-ADH 25 365 19.5 −175 9.00E−17 320 DHQ_synthase 37 346 −77.9 −116.5 1.00E−06 321 AlaDh_PNT_N 4 135 277 −6.3 4.20E−80 321 AlaDh_PNT_C 147 302 315.9 −13.4 8.10E−92 321 Shikimate_DH 152 276 2 −4.2 0.00013 322 Thiolase_N 14 277 421.7 −110.9 1.20E−123 322 Thiolase_C 284 406 219.2 −5.3 1.10E−62 323 Thiolase_N 14 277 452 −110.9 8.90E−133 323 Thiolase_C 284 406 218.6 −5.3 1.70E−62 324 SOUL 1 150 167.1 30.2 5.20E−47 325 AA_kinase 98 373 206.1 −39 9.60E−59 326 AA_kinase 10 285 196.9 −39 5.70E−56 327 AA_kinase 98 374 191.3 78.7 2.80E−54 327 NAD_binding_3 552 687 164.9 4 2.40E−46 327 Homoserine_dh 695 891 378.1 59.2 1.60E−110 328 HLH 204 253 56.2 35.3 1.20E−13 329 HLH 204 253 56.2 35.3 1.20E−13 330 HLH 258 307 70 32.4 8.90E−18 331 HLH 258 307 70 32.4 8.90E−18 332 Thioredoxin 7 105 42 −14.2 2.40E−09 332 Glutaredoxin 169 233 94.3 24.9 4.20E−25 332 Glutaredoxin 298 362 101.9 24.9 2.10E−27 332 Glutaredoxin 405 469 94 24.9 5.10E−25 334 Pkinase 143 405 308.2 154.7 1.80E−89 334 Pkinase_Tyr 143 443 −10.3 −149.1 4.90E−13 335 Gln-synt_N 17 97 146.5 13.5 8.20E−41 335 Gln-synt_C 103 355 291.7 −123.7 1.60E−84 336 Pkinase 143 405 308.2 154.7 1.80E−89 336 Pkinase_Tyr 143 443 −10.3 −149.1 4.90E−13 337 MethyltransfD12 23 264 358.3 −38.5 1.40E−104 338 bZIP_1 87 150 55.9 25.5 1.60E−13 338 bZIP_2 87 141 42 29.7 2.40E−09 338 MethyltransfD12 191 432 360.2 −38.5 4.00E−105 339 zf-B_box 1 47 29.8 22.6 1.10E−05 339 MethyltransfD12 248 489 358.3 −38.5 1.40E−104 340 TPP_enzyme_N 2 177 156.7 −34.7 7.20E−44 340 TPP_enzyme_M 193 325 89 −7.8 1.70E−23 340 TPP_enzyme_C 380 517 104.2 19.9 4.40E−28 341 RolB_RolC 11 197 186.6 109.2 6.90E−53 341 Amino_oxidase 246 729 336.5 57.4 5.10E−98 342 Amino_oxidase 55 532 117.8 45.7 3.60E−32 343 NIR_SIR_ferr 131 201 97.3 6.9 5.50E−26 343 NIR_SIR 209 369 171.8 −18.6 2.00E−48 343 NIR_SIR_ferr 390 459 95 6.9 2.50E−25 345 DEAD 121 290 239.3 67.8 9.80E−69 345 Helicase_C 359 435 120.7 96.2 4.90E−33 347 LRRNT_2 38 79 46 19.6 1.40E−10 347 LRR_1 108 130 15.4 11 0.24 347 LRR_1 132 154 14.8 11 0.35 347 LRR_1 156 178 12.2 11 1.4 347 LRR_1 180 201 13.7 11 0.73 347 LRR_1 228 250 11.2 11 2.2 347 LRR_1 252 274 14.6 11 0.42 347 LRR_1 299 321 12.7 11 1.1 347 LRR_1 347 369 16.7 11 0.1 347 LRR_1 419 441 12.8 11 1.1 347 LRR_1 443 465 15.3 11 0.25 347 LRR_1 467 489 14.6 11 0.4 347 LRR_1 491 513 16.4 11 0.12 347 LRR_1 539 560 16.7 11 0.094 347 Pkinase 662 927 117.1 90.1 6.00E−32 347 Pkinase_Tyr 662 927 74.3 31.1 4.50E−19 348 Homeobox 118 172 62.6 36.3 1.50E−15 348 HALZ 173 217 45.3 38.9 2.40E−10 349 Homeobox 139 193 62.6 36.3 1.50E−15 349 HALZ 194 238 45.3 38.9 2.40E−10 350 WD40 560 599 34.4 22.4 4.50E−07 350 WD40 646 683 30.5 22.4 6.80E−06 351 C4dic_mal_tran 199 465 224.8 −17.4 2.30E−64 352 Peroxidase 85 320 359.4 241.3 6.80E−105 353 Peroxidase 85 320 359.4 241.3 6.80E−105 354 LisH 7 33 40.7 29.6 5.80E−09 354 WD40 185 223 46.5 23.7 1.10E−10 354 WD40 244 282 30.2 23.7 8.60E−06 354 WD40 285 323 40.2 23.7 8.40E−09 354 WD40 327 364 23.5 23.5 0.00089 354 WD40 368 406 50.7 23.7 5.80E−12 354 WD40 410 457 46.3 23.7 1.20E−10 354 WD40 461 501 33.6 23.7 8.20E−07 355 bZIP_2 325 379 26.7 26.7 9.40E−05 355 bZIP_1 334 386 28.8 28.8 2.20E−05 356 FAD_binding_4 107 242 132.8 15.5 1.10E−36 356 ALO 266 581 −44 −93.7 1.40E−07 357 NAM 69 200 315.8 262 8.70E−92 358 Ribosomal_L21p 136 232 164.5 45.4 3.20E−46 359 BTB 192 301 85.6 19.5 1.70E−22 360 RWP-RK 104 155 106.6 89.8 8.40E−29 360 PB1 325 408 83.2 66.1 9.20E−22 361 Ammonium_transp 29 451 263 −105.1 7.00E−76 362 BTB 192 301 85.6 19.5 1.70E−22 363 OPT 85 744 815.2 −141.6 4.10E−242 364 DUF506 14 238 260.4 45.9 4.30E−75 366 FTCD_N 10 202 184 −99.6 4.10E−52 367 FTCD_N 7 199 231.8 −99.6 1.70E−66 368 SSF 44 461 −0.7 −114.1 8.50E−09 369 Aminotran_5 68 433 435.7 −139.8 7.30E−128 369 Beta_elim_lyase 93 373 −78.8 −110.3 5.00E−05 370 ABC_tran 67 262 118.5 9.8 2.10E−32 371 Monooxygenase_B 39 418 1037 175.6 0 372 PMEI 34 191 206.1 40.3 9.40E−59 374 CCT 141 185 83.6 63 7.20E−22 375 PHD 82 131 38.5 27 2.70E−08 375 SET 361 496 154.8 52.6 2.60E−43 376 AP2 111 163 73.7 56 6.70E−19 377 bZIP_2 390 444 33.6 21.1 8.20E−07 377 bZIP_1 392 453 32.9 26.8 1.30E−06 378 AP2 24 75 79.3 57 1.30E−20 379 A_thal_3526 17 70 119.3 100.1 1.30E−32 380 AP2 24 75 78.9 57 1.80E−20 381 Clp_N 17 67 39.1 1.9 1.70E−08 381 Clp_N 96 146 43.1 1.9 1.10E−09 381 AAA 202 396 46 14.1 1.50E−10 381 AAA_2 597 762 361.4 28 1.70E−105 381 AAA_5 601 767 15.6 4.1 4.80E−05 381 ClpB_D2-small 768 857 122.4 27.9 1.50E−33 382 eRF1_1 1 131 178 0.7 2.60E−50 382 eRF1_2 136 268 172.7 4.1 1.00E−48 382 eRF1_3 271 370 159.5 8.8 1.00E−44 387 zf-CCCH 113 138 31 2.1 4.80E−06 388 zf-B_box 1 47 42.6 15.5 1.50E−09 388 zf-B_box 48 90 24.8 15.5 0.00035 388 CCT 350 394 81.3 60.2 3.50E−21 389 HLH 128 178 33.4 10.8 9.20E−07 390 Arginosuc_synth 100 446 574.6 −233.5 1.10E−169 391 SH3BGR 204 301 −7.5 −5.5 0.006 391 Glutaredoxin 205 279 45.1 21 2.70E−10 392 AsnA 3 246 612.2 27.6 5.50E−181 393 NAD_binding_1 221 337 150.2 19.5 6.20E−42 394 cNMP_binding 39 131 74.2 21.2 4.70E−19 394 Crp 186 217 49.9 20.8 1.00E−11 395 PLAC8 71 166 175.3 63.2 1.80E−49 396 HSP20 42 148 109.2 34.1 1.40E−29 397 Myb_DNA-binding 14 61 44.2 28.5 5.30E−10 397 Myb_DNA-binding 67 112 47.4 28.5 5.60E−11 398 ATP-sulfurylase 136 462 529.7 −150.4 3.50E−156 399 WRKY 300 360 135.4 52.5 1.80E−37 400 Asp 106 439 −70.2 −149.4 2.10E−09 401 Gln-synt_N 42 122 26.1 14.1 4.70E−06 401 Gln-synt_C 141 444 −72.8 −123.9 2.90E−06 402 GATase_2 27 397 686.8 −57.2 1.90E−203 402 Glu_syn_central 477 773 530.8 32.1 1.70E−156 402 Glu_synthase 830 1221 802.2 −154.6 3.30E−238 402 GXGXG 1298 1493 323.1 9.5 5.60E−94 403 Form_Nir_trans 87 343 364.7 −118.6 1.70E−106 406 NAD_binding_2 51 213 202.4 12.2 1.20E−57 407 NAD_binding_2 1 162 189.3 12.2 1.00E−53 408 Glutaredoxin 39 103 76.8 30 7.70E−20 409 Glutaredoxin 59 123 111.1 55.5 3.80E−30 410 Homeobox 139 193 62.6 36.3 1.50E−15 410 HALZ 194 238 45.3 38.9 2.40E−10 411 Homeobox 137 193 59.7 36.3 1.10E−14 411 HALZ 194 238 45.3 38.9 2.40E−10 412 Homeobox 139 193 62.6 36.3 1.50E−15 412 HALZ 194 238 45.3 38.9 2.40E−10 414 CCT 228 272 90.1 74.2 7.80E−24 415 MBD 25 144 62.2 36 2.00E−15 416 MBD 21 94 49.3 13 1.50E−11 417 Pkinase 12 263 298.2 182.4 1.70E−86 417 Pkinase_Tyr 12 261 17.9 −122.6 6.50E−15 418 SRF-TF 3 53 85.4 21.1 2.00E−22 419 SRF-TF 11 65 30.5 18.7 6.90E−06 420 eIF-5a 85 154 129 10.6 1.50E−35 421 Homeobox 33 89 78.7 45.4 2.00E−20 421 START 236 457 196.1 6.7 9.50E−56 422 Homeobox 33 89 78.7 45.4 2.00E−20 422 START 236 457 196.1 6.7 9.50E−56 423 Homeobox 33 89 78.7 45.4 2.00E−20 423 START 236 457 196.1 6.7 9.50E−56 424 Homeobox 33 89 78.7 45.4 2.00E−20 424 START 236 457 196.1 6.7 9.50E−56 425 Arginase 61 336 283.6 0 4.40E−82 427 CBFB_NFYA 147 203 134.7 100.3 3.00E−37 428 CBFB_NFYA 148 204 139.4 89.5 1.10E−38 429 CBFB_NFYA 93 149 140.9 87.3 4.10E−39 430 CBFB_NFYA 70 126 140.2 89.5 6.60E−39 431 CBFB_NFYA 73 129 141.1 74.2 3.30E−39 432 PAS_2 103 219 243 82 7.50E−70 432 GAF 252 433 108.7 62.8 1.90E−29 432 Phytochrome 444 623 410.7 29 2.40E−120 432 PAS 654 770 105.5 8.3 1.80E−28 432 PAS 785 907 113.5 8.3 6.80E−31 432 HisKA 927 991 52.6 25.1 1.50E−12 432 HATPase_c 1039 1152 69.6 37.6 1.20E−17 433 HMG_box 44 113 115.8 37.7 1.40E−31 434 SPRY 77 224 25.9 22.2 1.70E−05 435 SPRY 77 224 25.9 22.2 1.70E−05 436 RRM_1 4 68 69.7 29.9 1.00E−17 436 RRM_1 97 160 52.9 29.9 1.30E−12 437 RRM_1 4 68 69.7 29.9 1.00E−17 437 RRM_1 97 160 52.9 29.9 1.30E−12 438 RRM_1 4 69 56.2 51.6 1.20E−13 438 zf-CCHC 85 102 31.4 18.3 3.10E−06 439 RRM_1 4 69 56.2 51.6 1.20E−13 439 zf-CCHC 85 102 31.4 18.3 3.10E−06 440 RRM_1 4 69 57.1 49.8 6.80E−14 440 zf-CCHC 86 103 33 18.3 1.20E−06 441 RRM_1 4 69 57.1 49.8 6.80E−14 441 zf-CCHC 86 103 33 18.3 1.20E−06 442 MMR_HSR1 119 240 131.3 66.5 3.10E−36 442 KH_2 345 409 87.5 25.3 4.60E−23 443 MMR_HSR1 119 240 131.3 66.5 3.10E−36 443 KH_2 345 409 87.5 25.3 4.60E−23 444 MMR_HSR1 119 240 131.3 66.5 3.10E−36 444 KH_2 345 409 87.5 25.3 4.60E−23 445 zf-C3HC4 152 193 40.2 29.7 8.40E−09 446 zf-C3HC4 152 193 40.2 29.7 8.40E−09 447 zf-C3HC4 152 193 40.2 29.7 8.40E−09 448 zf-C3HC4 152 193 40.2 29.7 8.40E−09 450 2-Hacid_dh 8 324 145.9 14 1.20E−40 450 2-Hacid_dh_C 113 292 276.4 117.8 6.30E−80 451 AlaDh_PNT_N 4 136 272.7 −6.3 8.50E−79 451 AlaDh_PNT_C 148 303 309.5 −13.4 7.00E−90 452 AlaDh_PNT_N 4 136 272.7 −6.3 8.50E−79 452 AlaDh_PNT_C 148 303 309.5 −13.4 7.00E−90 453 tRNA_synt_1c_R1 14 171 220.2 40.2 5.30E−63 453 tRNA_synt_1c_R2 172 264 109.8 27.2 9.20E−30 453 tRNA-synt_1c 271 577 565.2 −112.8 7.30E−167 453 tRNA-synt_1c_C 579 770 242.5 25.4 1.10E−69 454 SBP_bac_3 75 296 114.2 56.9 4.30E−31 455 Gln-synt_N 17 97 146.5 13.5 8.20E−41 455 Gln-synt_C 103 355 291.7 −123.7 1.60E−84 456 bZIP_1 76 139 43.5 27.4 8.20E−10 456 bZIP_2 76 130 42.5 27 1.70E−09 457 bZIP_1 76 139 43.5 27.4 8.20E−10 457 bZIP_2 76 130 42.5 27 1.70E−09 458 BPD_transp_2 5 354 254.5 33.4 2.60E−73 459 BPD_transp_2 82 431 254.5 33.4 2.60E−73 460 Asp_decarbox 2 119 198.7 156.7 1.60E−56 462 Glt_symporter 4 370 805.4 −247.2 3.70E−239 463 Glt_symporter 4 370 805.4 −247.2 3.70E−239 464 SBP_bac_3 75 296 114.2 56.9 4.30E−31 465 SBP_bac_3 75 296 114.2 56.9 4.30E−31 467 AlaDh_PNT_N 4 136 262.3 −6.3 1.20E−75 467 AlaDh_PNT_C 148 302 275.9 −13.4 9.10E−80 468 Nitroreductase 20 181 82.5 37 1.60E−21 469 Acid_phosphat_B 1 179 89.8 −18.9 9.70E−24 470 bZIP_1 76 139 43.5 27.4 8.20E−10 470 bZIP_2 76 130 42.5 27 1.70E−09 471 Ubiquitin 22 92 45.4 19.6 2.20E−10 472 LIM 172 228 29 9.4 2.50E−06 473 LIM 147 203 42 2.4 2.40E−09 474 zf-C3HC4 197 237 43.5 21.9 8.40E−10 475 RCC1 95 145 24.5 21.5 0.00045 475 RCC1 266 315 25.8 21.5 0.00018 475 RCC1 350 398 48.9 21.5 2.00E−11 476 Sulfate_transp 198 510 494.9 −44.7 1.10E−145 476 STAS 533 652 84.7 1.5 3.30E−22 477 RCC1 95 145 24.5 21.5 0.00045 477 RCC1 266 315 25.8 21.5 0.00018 477 RCC1 350 398 48.9 21.5 2.00E−11 479 Glyco_transf_20 58 544 840.5 −201.3 9.90E−250 479 Trehalose_PPase 593 829 338.3 −32.2 1.50E−98 480 RRM_1 19 89 25 18 0.0003 481 Asp 89 438 −88.1 −149.4 2.40E−08 482 Ank 44 81 12 1.2 0.67 482 Ank 83 112 21.9 1.2 0.0026 482 Ank 118 150 16.3 1.2 0.13 482 Ank 152 184 17.2 1.2 0.07 482 Ank 186 218 31.4 1.2 3.70E−06 482 Ank 220 252 24.2 1.2 0.00055 482 Ank 254 287 29 1.2 2.00E−05 482 Ank 288 320 1.6 1.2 22 484 DSPc 105 240 25.3 −4.2 1.90E−06 485 Usp 8 162 86.9 26.5 7.20E−23 486 U-box 17 94 76.1 33.2 1.30E−19 487 U-box 10 84 83.9 33.2 5.70E−22 493 MIT 5 73 103.3 38.4 8.20E−28 493 AAA 169 353 231.7 43.1 1.80E−66 493 Vps4_C 368 433 127.3 −2.2 5.10E−35 494 AP2 80 132 77.7 59.9 4.20E−20 495 Ssl1 37 292 530.7 −115.9 1.80E−156 495 C1_4 374 422 98.3 50.7 2.60E−26 496 POX 213 346 −18.9 −23 2.80E−05 496 Homeobox 396 454 −2.1 −3.7 0.019 497 POX 214 350 −0.6 −24.9 6.60E−07 497 Homeobox 404 462 −2.5 −3.7 0.021 498 Dimerisation 34 82 71.7 35.1 2.70E−18 498 Methyltransf_2 91 342 278.1 −95.8 2.00E−80 499 DUF640 1 160 251 172 2.90E−72 500 p450 2 479 214.1 −29.9 3.80E−61 501 MFS_1 47 528 24.1 24.1 0.00057 501 PTR2 109 520 323.9 −39.5 3.30E−94 502 Pkinase 16 269 336.6 200.3 4.70E−98 502 Pkinase_Tyr 16 267 36.2 −68.3 3.90E−16 502 NAF 311 372 112.2 25.9 1.70E−30 503 Acyl-ACP_TE 92 367 438 40.9 1.40E−128 504 PAS_2 69 185 230.5 48 4.20E−66 504 GAF 218 402 111.1 45.8 3.60E−30 504 Phytochrome 413 592 421.7 29 1.20E−123 504 PAS 620 735 101.9 8.3 2.20E−27 504 PAS_4 626 740 35.5 16.4 2.10E−07 504 PAS 750 875 100 8.3 8.30E−27 504 HisKA 895 959 44.5 22.5 4.10E−10 504 HATPase_c 1007 1118 72.8 37.5 1.20E−18 505 PAS_2 69 185 230.5 48 4.20E−66 505 GAF 218 402 108 45.8 3.20E−29 505 Phytochrome 413 592 421.7 29 1.20E−123 505 PAS 620 735 101.9 8.3 2.20E−27 505 PAS_4 626 740 35.5 16.4 2.10E−07 505 PAS 750 875 100 8.3 8.30E−27 505 HisKA 895 959 44.5 22.5 4.10E−10 505 HATPase_c 1007 1118 72.8 37.5 1.20E−18 506 PAS_2 69 185 230.5 48 4.20E−66 506 GAF 218 402 105.4 45.8 1.90E−28 506 Phytochrome 413 592 421.7 29 1.20E−123 506 PAS 620 735 101.9 8.3 2.20E−27 506 PAS_4 626 740 35.5 16.4 2.10E−07 506 PAS 750 875 100 8.3 8.30E−27 506 HisKA 895 959 44.5 22.5 4.10E−10 506 HATPase_c 1007 1118 72.8 37.5 1.20E−18 507 zf-C3HC4 152 193 40.2 29.7 8.40E−09 508 zf-C3HC4 152 193 40.2 29.7 8.40E−09 509 CSD 4 70 156.9 6.7 6.20E−44 510 Pkinase 17 329 223.2 116.5 6.80E−64 510 Pkinase_Tyr 17 329 −72.7 −149.3 6.80E−09 511 SIS 56 193 64.5 12.1 4.00E−16 511 CBS 226 347 78.5 25.8 2.50E−20 513 BCCT 15 504 945.1 −372.2 3.30E−281 514 Dehydrin 1 93 40.4 23 7.30E−09 515 Nitroreductase 20 181 82.5 37 1.60E−21 516 Auxin_inducible 29 95 139.1 43.4 1.40E−38 517 zf-CCHC 602 619 19.6 19.6 0.00034 517 Plus-3 699 804 138.8 52.5 1.70E−38 518 U-box 7 83 86.3 36.7 1.10E−22 519 bZIP_1 273 333 45.2 30.3 2.50E−10 519 bZIP_2 273 327 47.2 32.8 6.40E−11 520 FAD_binding_3 80 438 −92.9 −134.3 3.00E−06 520 FHA 558 634 61.7 36.3 2.80E−15 521 LIM 10 66 63.1 36.4 1.00E−15 521 LIM 109 165 80 36.4 8.40E−21 522 Ndr 23 307 507.9 −78.4 1.30E−149 523 Pkinase 81 374 221.1 100.3 2.80E−63 523 Pkinase_Tyr 81 374 −78.4 −126.6 1.60E−08 524 SRF-TF 9 59 121.8 12.8 2.30E−33 524 K-box 75 174 156.5 74.3 7.80E−44 525 Sina 98 297 406.3 16.2 5.10E−119 526 HLH 28 78 10.6 9 0.013 527 Myb_DNA-binding 10 57 44.7 29.4 3.50E−10 527 Myb_DNA-binding 63 108 52.8 29.4 1.30E−12 528 HLH 61 144 22.1 22.1 0.00078 529 DUF828 85 273 300.8 11.5 2.80E−87 529 PH_2 290 397 32.2 −0.7 7.90E−07 530 SRF-TF 9 59 115.9 14.8 1.30E−31 530 K-box 74 173 138.8 5.9 1.70E−38 531 PBP 20 165 220.2 −19.8 5.20E−63 532 PTR2 97 499 485.3 16.2 8.10E−143 533 ABC_membrane 71 350 216 33.5 1.00E−61 533 ABC_tran 424 609 222.2 71.8 1.30E−63 534 GATase_2 2 173 −17.6 −89.2 1.00E−07 534 Asn_synthase 227 460 297.7 −50.9 2.60E−86 535 Acetate_kinase 25 420 605.3 −200.3 6.30E−179 536 PTA_PTB 452 770 511.1 −121.8 1.50E−150 537 OKR_DC_1 20 491 143 −45.4 9.50E−40 537 OKR_DC_1_C 485 584 −4.2 −51.6 1.60E−07 538 Mito_carr 79 176 131.9 23.8 2.10E−36 538 Mito_carr 184 281 107.7 23.8 4.00E−29 538 Mito_carr 285 375 88.8 23.8 1.90E−23 539 AP2 114 165 76 27 1.40E−19 542 60KD_IMP 128 347 360.6 7.8 2.90E−105 545 eIF-5a 86 155 133.7 17.4 6.00E−37 546 Aa_trans 32 463 484.6 −127.8 1.40E−142 547 zf-B_box 1 45 38.3 36.1 3.10E−08 548 MFS_1 47 528 24.1 24.1 0.00057 548 PTR2 109 520 323.9 −39.5 3.30E−94 549 Homeobox 17 71 62.8 −2.9 1.30E−15 549 HALZ 72 116 73.7 20 6.80E−19 550 Homeobox 68 122 77 12.5 6.70E−20 550 HALZ 123 167 81.9 19.5 2.30E−21 551 Homeobox 68 122 77 12.5 6.70E−20 551 HALZ 123 167 81.9 19.5 2.30E−21 552 Homeobox 68 122 77 12.5 6.70E−20 552 HALZ 123 167 81.9 19.5 2.30E−21 553 Homeobox 71 125 75.9 52.9 1.40E−19 553 HALZ 126 170 38.4 19.5 2.90E−08 554 PP2C 35 319 82.4 44 1.60E−21 555 PP2C 46 328 119.2 44 1.40E−32 556 PP2C 40 319 112.1 −25.3 1.90E−30 557 Response_reg 13 125 87.6 5.1 4.60E−23 557 Myb_DNA-binding 202 252 45.8 22.8 1.70E−10 558 AP2 151 203 84.3 58.7 4.30E−22 560 F-box 51 106 29.6 15.3 1.30E−05 560 Tub 117 442 535 81.2 9.00E−158 561 AA_permease 78 509 33.9 −118 6.60E−07 562 AA_permease 65 510 41.7 −118 2.90E−09 563 Transaldolase 39 258 17.3 −12.6 6.50E−10 564 Acetate_kinase 25 420 605.3 −200.3 6.30E−179 565 OKR_DC_1 20 491 143 −45.4 9.50E−40 565 OKR_DC_1_C 485 584 −4.2 −51.6 1.60E−07 566 Gal_Lectin 75 152 66.5 27 1.00E−16 567 zf-C3HC4 107 148 32.2 20.4 2.10E−06 568 Aa_trans 32 429 241.2 −100.4 2.60E−69 569 zf-C3HC4 85 135 36.6 23.4 9.90E−08 570 HR_lesion 1 138 322 103.1 1.20E−93 571 zf-Dof 43 105 140.8 27.7 4.30E−39 572 zf-Dof 43 105 140.8 27.7 4.30E−39 573 zf-Dof 43 105 140.8 27.7 4.30E−39 574 zf-B_box 1 46 30.1 22.6 9.20E−06 575 PAS_2 69 185 230.5 48 4.20E−66 575 GAF 218 402 111.1 45.8 3.60E−30 575 Phytochrome 413 592 421.7 29 1.20E−123 575 PAS 620 735 101.9 8.3 2.20E−27 575 PAS_4 626 740 35.5 16.4 2.10E−07 575 PAS 750 875 100 8.3 8.30E−27 575 HisKA 895 959 44.5 22.5 4.10E−10 575 HATPase_c 1007 1118 72.8 35.3 1.20E−18 576 PAS_2 69 185 230.5 48 4.20E−66 576 GAF 218 402 111.1 45.8 3.60E−30 576 Phytochrome 413 592 421.7 29 1.20E−123 576 PAS 620 735 101.9 8.3 2.20E−27 576 PAS_4 626 740 35.5 16.4 2.10E−07 576 PAS 750 875 100 8.3 8.30E−27 576 HisKA 895 959 44.5 22.5 4.10E−10 576 HATPase_c 1007 1118 72.8 35.3 1.20E−18 577 PAS_2 69 185 230.5 48 4.20E−66 577 GAF 218 402 111.1 45.8 3.60E−30 577 Phytochrome 413 592 421.7 29 1.20E−123 577 PAS 620 735 101.9 8.3 2.20E−27 577 PAS_4 626 740 35.5 16.4 2.10E−07 577 PAS 750 875 100 8.3 8.30E−27 577 HisKA 895 959 44.5 22.5 4.10E−10 577 HATPase_c 1007 1118 72.8 35.3 1.20E−18 578 Ammonium_transp 47 470 651.5 −140.2 8.00E−193 580 Thg1 1 255 283.3 28.8 5.50E−82 580 Thg1 273 520 352.9 28.8 6.10E−103 581 DUF647 177 593 868.5 29.4 3.80E−258 582 AP2 95 147 79.8 52.3 1.00E−20 584 Peptidase_C2 1696 1997 285.6 −156.6 1.10E−82 584 Calpain_III 1999 2149 −9.8 −54.1 3.00E−06 585 HSP70 37 644 1324.4 412 0 586 Pkinase 4 260 305.1 111.6 1.50E−88 586 Pkinase_Tyr 4 264 −38.8 −125 3.80E−11 587 WRKY 165 223 131.8 5.7 2.30E−36 587 WRKY 327 386 148.1 5.7 2.70E−41 588 WRKY 183 241 144.9 5.7 2.60E−40 588 WRKY 361 420 151.5 5.7 2.60E−42 589 LysM 103 148 20.7 20.1 0.0039 590 LRRNT_2 357 400 30.5 18.9 6.80E−06 590 LRR_1 431 452 11.8 10.9 1.7 590 Pkinase_Tyr 587 867 120.2 −144.1 6.70E−33 590 Pkinase 595 867 114.7 74.8 3.00E−31 591 GATase_2 2 161 78.2 −89.2 2.90E−20 591 Asn_synthase 209 450 337.9 −46 2.00E−98 592 GH3 31 592 1167.9 −249.6 0 593 MFS_1 37 502 38.5 23.9 2.70E−08 593 PTR2 99 494 362.6 91.2 7.30E−106 594 Glutaredoxin 89 153 85.3 17.5 2.20E−22 595 Glutaredoxin 30 94 97.7 55.5 3.90E−26 595 Glutaredoxin 166 230 90.6 55.5 5.30E−24 595 Glutaredoxin 273 337 101.9 55.5 2.10E−27 596 Glutaredoxin 13 75 59 39.4 1.70E−14 597 GTP1_OBG 212 377 230.5 −51.4 4.30E−66 597 MMR_HSR1 379 502 136.2 38 1.10E−37 597 DUF1967 585 654 104.9 10.4 2.80E−28 598 HLH 70 120 39.8 10.5 1.10E−08 600 LRR_1 116 138 14.5 11.1 0.44 600 LRR_1 140 162 12.9 11.1 1 600 LRR_1 164 186 15.2 11.1 0.27 600 LRR_1 188 209 13.7 11.1 0.73 600 Pkinase_Tyr 497 764 39 −37.1 2.50E−16 601 DEAD_2 72 256 297.9 29.3 2.20E−86 601 DUF1227 267 413 296.6 165.1 5.20E−86 602 NUDIX 32 173 60.1 2.2 8.60E−15 604 zf-C3HC4 27 67 31.5 19.1 3.40E−06 605 SRF-TF 9 59 116 14.8 1.20E−31 605 K-box 75 176 99.8 5.9 9.20E−27 606 SRF-TF 9 59 109.5 14.8 1.10E−29 606 K-box 74 173 96.9 5.9 7.20E−26 607 Aa_trans 58 464 166.2 133 9.40E−47 608 Aa_trans 26 463 373 −127.8 5.50E−109 612 Myb_DNA-binding 20 67 46.2 26.4 1.30E−10 612 Myb_DNA-binding 73 118 44.6 26.4 3.80E−10 613 Globin 14 123 64.6 −6.9 3.80E−16 613 FAD_binding_6 177 276 42.5 −2.2 1.60E−09 613 NAD_binding_1 287 396 88.4 −1.7 2.60E−23 614 Globin 6 103 61.7 9.9 2.80E−15 614 FAD_binding_6 156 262 42.7 −4.5 1.40E−09 614 NAD_binding_1 271 379 66.1 −3.4 1.30E−16

TABLE 10 Pfam module annotation PEP SEQ ID NO Pfam domain module Position 308 Aminotran_1_2 69-470 309 Homeobox::HALZ 65-119::120-164 310 bZIP_2 108-162 312 Peptidase_S10 39-504 313 SBP 103-181 315 GDPD 436-728 317 DUF1292 111-180 318 Aminotran_1_2 127-473 319 FAD_binding_4::Lact-deh- 50-191::279-570 memb 320 Fe-ADH 25-365 321 AlaDh_PNT_N::AlaDh_PNT_C 4-135::147-302 322 Thiolase_N::Thiolase_C 14-277::284-406 323 Thiolase_N::Thiolase_C 14-277::284-406 324 SOUL 1-150 325 AA_kinase 98-373 326 AA_kinase 10-285 327 AA_kinase::NAD_binding_3:: 98-374::552-687::695-891 Homoserine_dh 328 HLH 204-253 329 HLH 204-253 330 HLH 258-307 331 HLH 258-307 332 Thioredoxin::Glutaredoxin::Glutaredoxin:: 7-105::169-233::298-362:: Glutaredoxin 405-469 334 Pkinase 143-405 335 Gln-synt_N::Gln-synt_C 17-97::103-355 336 Pkinase 143-405 337 MethyltransfD12 23-264 338 bZIP_1::MethyltransfD12 87-150::191-432 339 zf-B_box::MethyltransfD12 1-47::248-489 340 TPP_enzyme_N::TPP_enzyme_M:: 2-177::193-325::380-517 TPP_enzyme_C 341 RolB_RolC::Amino_oxidase 11-197::246-729 342 Amino_oxidase 55-532 343 NIR_SIR_ferr::NIR_SIR::NIR_SIR_ferr 131-201::209-369::390-459 345 DEAD::Helicase_C 121-290::359-435 347 LRRNT_2::LRR_1::LRR_1::LRR_1:: 38-79::108-130::132-154:: LRR_1::LRR_1::LRR_1:: 156-178::180-201::228-250:: LRR_1::LRR_1::LRR_1::LRR_1:: 252-274::299-321::347-369:: LRR_1::LRR_1::LRR_1:: 419-441::443-465::467-489:: Pkinase 491-513::539-560::662-927 348 Homeobox::HALZ 118-172::173-217 349 Homeobox::HALZ 139-193::194-238 350 WD40::WD40 560-599::646-683 351 C4dic_mal_tran 199-465 352 Peroxidase 85-320 353 Peroxidase 85-320 354 LisH::WD40::WD40::WD40:: 7-33::185-223::244-282::285-323:: WD40::WD40::WD40::WD40 327-364::368-406::410-457:: 461-501 355 bZIP_2::bZIP_1 325-379::334-386 356 FAD_binding_4::ALO 107-242::266-581 357 NAM 69-200 358 Ribosomal_L21p 136-232 359 BTB 192-301 360 RWP-RK::PB1 104-155::325-408 361 Ammonium_transp 29-451 362 BTB 192-301 363 OPT 85-744 364 DUF506 14-238 366 FTCD_N 10-202 367 FTCD_N 7-199 368 SSF 44-461 369 Aminotran_5 68-433 370 ABC_tran 67-262 371 Monooxygenase_B 39-418 372 PMEI 34-191 374 CCT 141-185 375 PHD::SET 82-131::361-496 376 AP2 111-163 377 bZIP_2 390-444 378 AP2 24-75 379 A_thal_3526 17-70 380 AP2 24-75 381 Clp_N::Clp_N::AAA::AAA_2:: 17-67::96-146::202-396::597-762:: ClpB_D2-small 768-857 382 eRF1_1::eRF1_2::eRF1_3 1-131::136-268::271-370 387 zf-CCCH 113-138 388 zf-B_box::zf-B_box::CCT 1-47::48-90::350-394 389 HLH 128-178 390 Arginosuc_synth 100-446 391 Glutaredoxin 205-279 392 AsnA 3-246 392 NAD_binding_1 221-337 394 cNMP_binding::Crp 39-131::186-217 395 PLAC8 71-166 396 HSP20 42-148 397 Myb_DNA- 14-61::67-112 binding::Myb_DNA-binding 398 ATP-sulfurylase 136-462 399 WRKY 300-360 400 Asp 106-439 401 Gln-synt_N::Gln-synt_C 42-122::141-444 402 GATase_2::Glu_syn_central::Glu_synthase:: 27-397::477-773::830-1221:: GXGXG 1298-1493 403 Form_Nir_trans 87-343 406 NAD_binding_2 51-213 407 NAD_binding_2 1-162 408 Glutaredoxin 39-103 409 Glutaredoxin 59-123 410 Homeobox::HALZ 139-193::194-238 411 Homeobox::HALZ 137-193::194-238 412 Homeobox::HALZ 139-193::194-238 414 CCT 228-272 415 MBD 25-144 416 MBD 21-94 417 Pkinase 12-263 418 SRF-TF March 1953 419 SRF-TF November 1965 420 eIF-5a 85-154 421 Homeobox::START 33-89::236-457 422 Homeobox::START 33-89::236-457 423 Homeobox::START 33-89::236-457 424 Homeobox::START 33-89::236-457 425 Arginase 61-336 427 CBFB_NFYA 147-203 428 CBFB_NFYA 148-204 429 CBFB_NFYA 93-149 430 CBFB_NFYA 70-126 431 CBFB_NFYA 73-129 432 PAS_2::GAF::Phytochrome::PAS:: 103-219::252-433::444-623:: PAS::HisKA::HATPase_c 654-770::785-907::927-991:: 1039-1152 433 HMG_box 44-113 434 SPRY 77-224 435 SPRY 77-224 436 RRM_1::RRM_1 4-68::97-160 437 RRM_1::RRM_1 4-68::97-160 438 RRM_1::zf-CCHC 4-69::85-102 439 RRM_1::zf-CCHC 4-69::85-102 440 RRM_1::zf-CCHC 4-69::86-103 441 RRM_1::zf-CCHC 4-69::86-103 442 MMR_HSR1::KH_2 119-240::345-409 443 MMR_HSR1::KH_2 119-240::345-409 444 MMR_HSR1::KH_2 119-240::345-409 445 zf-C3HC4 152-193 446 zf-C3HC4 152-193 447 zf-C3HC4 152-193 448 zf-C3HC4 152-193 450 2-Hacid_dh_C 113-292 451 AlaDh_PNT_N::AlaDh_PNT_C 4-136::148-303 452 AlaDh_PNT_N::AlaDh_PNT_C 4-136::148-303:: 453 tRNA_synt_1c_R1::tRNA_synt_1c_R2:: 14-171::172-264::271-577 tRNA-synt_1c::tRNA- 579-770 synt_1c_C 454 SBP_bac_3 75-296 455 Gln-synt_N::Gln-synt_C 17-97::103-355 456 bZIP_1 76-139 457 bZIP_1 76-139 458 BPD_transp_2 5-354 459 BPD_transp_2 82-431 460 Asp_decarbox 2-119 462 Glt_symporter 4-370 463 Glt_symporter 4-370 464 SBP_bac_3 75-296 465 SBP_bac_3 75-296 467 AlaDh_PNT_N::AlaDh_PNT_C 4-136::148-302 468 Nitroreductase 20-181 469 Acid_phosphat_B 1-179 470 bZIP_1 76-139 471 Ubiquitin 22-92 472 LIM 172-228 473 LIM 147-203 474 zf-C3HC4 197-237 475 RCC1::RCC1::RCC1 95-145::266-315::350-398 476 Sulfate_transp::STAS 198-510::533-652 477 RCC1::RCC1::RCC1 95-145::266-315::350-398 479 Glyco_transf_20::Trehalose_PPase 58-544::593-829 480 RRM_1 19-89 481 Asp 89-438 482 Ank::Ank::Ank::Ank::Ank::Ank:: 44-81::83-112::118-150::152-184:: Ank::Ank 186-218::220-252::254-287:: 288-320 484 DSPc 105-240 485 Usp 8-162 486 U-box 17-94 487 U-box October 1984 493 MIT::AAA::Vps4_C 5-73::169-353::368-433 494 AP2 80-132 495 Ssl1::C1_4 37-292::374-422 496 POX::Homeobox 213-346::396-454 497 POX::Homeobox 214-350::404-462 498 Dimerisation::Methyltransf_2 34-82::91-342 499 DUF640 1-160 500 p450 2-479 501 PTR2 109-520 502 Pkinase::NAF 16-269::311-372 503 Acyl-ACP_TE 92-367 504 PAS_2::GAF::Phytochrome::PAS:: 69-185::218-402::413-592:: PAS::HisKA::HATPase_c 620-735::750-875::895-959 1007-1118 505 PAS_2::GAF::Phytochrome::PAS:: 69-185::218-402::413-592:: PAS::HisKA::HATPase_c 620-735::750-875::895-959:: 1007-1118 506 PAS_2::GAF::Phytochrome::PAS:: 69-185::218-402::413-592:: PAS::HisKA::HATPase_c 620-735::750-875::895-959:: 1007-1118 507 zf-C3HC4 152-193 508 zf-C3HC4 152-193 509 CSD April 1970 510 Pkinase 17-329 511 SIS::CBS 56-193::226-347 513 BCCT 15-504 514 Dehydrin January 1993 515 Nitroreductase 20-181 516 Auxin_inducible 29-95 517 zf-CCHC::Plus-3 602-619::699-804 518 U-box July 1983 519 bZIP_2 273-327 520 FAD_binding_3::FHA 80-438::558-634 521 LIM::LIM 10-66::109-165 522 Ndr 23-307 523 Pkinase 81-374 524 SRF-TF::K-box 9-59::75-174 525 Sina 98-297 526 HLH 28-78 527 Myb_DNA- 10-57::63-108 binding::Myb_DNA-binding 528 HLH 61-144 529 DUF828::PH_2 85-273::290-397 530 SRF-TF::K-box 9-59::74-173 531 PBP 20-165 532 PTR2 97-499 533 ABC_membrane::ABC_tran 71-350::424-609 534 GATase_2::Asn_synthase 2-173::227-460 535 Acetate_kinase 25-420 536 PTA_PTB 452-770 537 OKR_DC_1::OKR_DC_1_C 20-491::485-584 538 Mito_carr::Mito_carr::Mito_carr 79-176::184-281::285-375 539 AP2 114-165 542 60KD_IMP 128-347 545 eIF-5a 86-155 546 Aa_trans 32-463 547 zf-B_box January 1945 548 PTR2 109-520 549 Homeobox::HALZ 17-71::72-116 550 Homeobox::HALZ 68-122::123-167 551 Homeobox::HALZ 68-122::123-167 552 Homeobox::HALZ 68-122::123-167 553 Homeobox::HALZ 71-125::126-170 554 PP2C 35-319 555 PP2C 46-328 556 PP2C 40-319 557 Response_reg::Myb_DNA- 13-125::202-252 binding 558 AP2 151-203 560 F-box::Tub 51-106::117-442 561 AA_permease 78-509 562 AA_permease 65-510 563 Transaldolase 39-258 564 Acetate_kinase 25-420 565 OKR_DC_1::OKR_DC_1_C 20-491::485-584 566 Gal_Lectin 75-152 567 zf-C3HC4 107-148 568 Aa_trans 32-429 569 zf-C3HC4 85-135 570 HR_lesion 1-138 571 zf-Dof 43-105 572 zf-Dof 43-105 573 zf-Dof 43-105 574 zf-B_box January 1946 575 PAS_2::GAF::Phytochrome::PAS:: 69-185::218-402::413-592:: PAS::HisKA::HATPase_c 620-735::750-875::895-959:: 1007-1118 576 PAS_2::GAF::Phytochrome::PAS:: 69-185::218-402::413-592:: PAS::HisKA::HATPase_c 620-735::750-875::895-959:: 1007-1118 577 PAS_2::GAF::Phytochrome::PAS:: 69-185::218-402::413-592:: PAS::HisKA::HATPase_c 620-735::750-875::895-959:: 1007-1118 578 Ammonium_transp 47-470 580 Thg1::Thg1 1-255::273-520 581 DUF647 177-593 582 AP2 95-147 584 Peptidase_C2::Calpain_III 1696-1997::1999-2149 585 HSP70 37-644 586 Pkinase 4-260 587 WRKY::WRKY 165-223::327-386 588 WRKY::WRKY 183-241::361-420 589 LysM 103-148 590 LRRNT_2::LRR_1::Pkinase_Tyr 357-400::431-452::587-867 591 GATase_2::Asn_synthase 2-161::209-450 592 GH3 31-592 593 PTR2 99-494 594 Glutaredoxin 89-153 595 Glutaredoxin::Glutaredoxin::Glutaredoxin 30-94::166-230::273-337 596 Glutaredoxin 13-75 597 GTP1_OBG::MMR_HSR1::DUF1967 212-377::379-502::585-654 598 HLH 70-120 600 LRR_1::LRR_1::LRR_1::LRR_1:: 116-138::140-162::164-186:: Pkinase_Tyr 188-209::497-764 601 DEAD_2::DUF1227 72-256::267-413 602 NUDIX 32-173 604 zf-C3HC4 27-67 605 SRF-TF::K-box 9-59::75-176 606 SRF-TF::K-box 9-59::74-173 607 Aa_trans 58-464 608 Aa_trans 26-463 612 Myb_DNA- 20-67::73-118 binding::Myb_DNA-binding 613 Globin::FAD_binding_6::NAD_binding_1 14-123::177-276::287-396 614 Globin::FAD_binding_6::NAD_binding_1 6-103::156-262::271-379

TABLE 11 Description of Pfam domains Accession Gathering Pfam domain name number cutoff Domain description 14-3-3 PF00244.12 25.0000; 14-3-3 protein 2-Hacid_dh PF00389.22 11.2000; D-isomer specific 2-hydroxyacid dehydrogenase, catalytic domain 2-Hacid_dh_C PF02826.11 −78.6000; D-isomer specific 2-hydroxyacid dehydrogenase, NAD binding domain 3-alpha PF03475.6 25.0000; 3-alpha domain 3HCDH_N PF02737.10 −78.7000; 3-hydroxyacyl-CoA dehydrogenase, NAD binding domain 60KD_IMP PF02096.12 −95.0000; 60 Kd inner membrane protein AAA PF00004.21 12.3000; ATPase family associated with various cellular activities (AAA) AAA_2 PF07724.6 −5.0000; ATPase family associated with various cellular activities (AAA) AAA_5 PF07728.6 4.0000; ATPase family associated with various cellular activities (AAA) AA_kinase PF00696.20 −40.0000; Amino acid kinase family AA_permease PF00324.13 −120.8000; Amino acid permease ABC_membrane PF00664.15 19.4000; ABC transporter transmembrane region ABC_membrane_2 PF06472.7 −94.9000; ABC transporter transmembrane region 2 ABC_tran PF00005.19 9.5000; ABC transporter ACP_syn_III_C PF08541.2 −20.5000; 3-Oxoacyl-[acyl-carrier-protein (ACP)] synthase III C terminal ACT PF01842.17 0.7000; ACT domain AFG1_ATPase PF03969.8 −177.5000; AFG1-like ATPase ALO PF04030.6 −99.7000; D-arabinono-1,4-lactone oxidase AMP-binding PF00501.20 0.0000; AMP-binding enzyme AP2 PF00847.12 21.7000; AP2 domain APH PF01636.15 −1.3000; Phosphotransferase enzyme family APS_kinase PF01583.12 25.0000; Adenylylsulphate kinase ATP-sulfurylase PF01747.9 −163.3000; ATP-sulfurylase A_thal_3526 PF09713.2 25.0000; Plant protein 1589 of unknown function (A_thal_3526) Aa_trans PF01490.10 −128.4000; Transmembrane amino acid transporter protein Abhydrolase_1 PF00561.12 10.3000; alpha/beta hydrolase fold Abi PF02517.8 −5.0000; CAAX amino terminal protease family Acetate_kinase PF00871.9 −206.2000; Acetokinase family Acetyltransf_1 PF00583.16 17.8000; Acetyltransferase (GNAT) family Acid_phosphat_B PF03767.6 −36.0000; HAD superfamily, subfamily IIIB (Acid phosphatase) Actin PF00022.11 −144.0000; Actin Acyl-ACP_TE PF01643.9 −106.0000; Acyl-ACP thioesterase AdoHcyase_NAD PF00670.13 −23.6000; S-adenosyl-L-homocysteine hydrolase, NAD binding domain AlaDh_PNT_C PF01262.13 −43.4000; Alanine dehydrogenase/PNT, C-terminal domain AlaDh_PNT_N PF05222.7 −11.7000; Alanine dehydrogenase/PNT, N-terminal domain Ala_racemase_N PF01168.12 −34.3000; Alanine racemase, N-terminal domain Alliinase_C PF04864.5 −233.5000; Allinase Amidase PF01425.13 −133.0000; Amidase Amidohydro_1 PF01979.12 −37.4000; Amidohydrolase family Amino_oxidase PF01593.16 −11.4000; Flavin containing amine oxidoreductase Aminotran_1_2 PF00155.13 −57.5000; Aminotransferase class I and II Aminotran_3 PF00202.13 −206.1000; Aminotransferase class-III Aminotran_5 PF00266.11 −164.4000; Aminotransferase class-V Ammonium_transp PF00909.13 −144.0000; Ammonium Transporter Family Ank PF00023.22 0.0000; Ankyrin repeat Anth_synt_I_N PF04715.5 25.0000; Anthranilate synthase component I, N terminal region ApbA PF02558.8 −16.2000; Ketopantoate reductase PanE/ApbA Arginase PF00491.13 −48.2000; Arginase family Arginosuc_synth PF00764.11 −236.3000; Arginosuccinate synthase Arm PF00514.15 17.0000; Armadillo/beta-catenin-like repeat AsnA PF03590.7 25.0000; Aspartate-ammonia ligase Asn_synthase PF00733.13 −52.8000; Asparagine synthase Asp PF00026.15 −153.8000; Eukaryotic aspartyl protease Asp_decarbox PF02261.8 −44.4000; Aspartate decarboxylase Auxin_inducible PF02519.6 25.0000; Auxin responsive protein BAG PF02179.8 25.0000; BAG domain BAH PF01426.10 7.0000; BAH domain BCCT PF02028.9 −386.1000; BCCT family transporter BNR PF02012.12 18.0000; BNR/Asp-box repeat BPD_transp_1 PF00528.14 −6.6000; Binding-protein-dependent transport system inner membrane component BPD_transp_2 PF02653.8 −66.1000; Branched-chain amino acid transport system/ permease component BSD PF03909.9 26.4000; BSD domain BTB PF00651.23 6.2000; BTB/POZ domain Bac_DnaA PF00308.10 −108.0000; Bacterial dnaA protein BenE PF03594.5 −283.2000; Benzoate membrane transport protein Beta_elim_lyase PF01212.13 −110.3000; Beta-eliminating lyase Big_2 PF02368.10 0.0000; Bacterial Ig-like domain (group 2) C1_1 PF00130.14 11.4000; Phorbol esters/diacylglycerol binding domain (C1 domain) C1_4 PF07975.4 25.0000; TFIIH C1-like domain C2 PF00168.22 3.7000; C2 domain C4dic_mal_tran PF03595.9 −43.6000; C4-dicarboxylate transporter/malic acid transport protein CBFB_NFYA PF02045.7 25.0000; CCAAT-binding transcription factor (CBF- B/NF-YA) subunit B CBS PF00571.20 19.5000; CBS domain pair CCT PF06203.6 25.0000; CCT motif CDC48_2 PF02933.9 25.0000; Cell division protein 48 (CDC48), domain 2 CDC48_N PF02359.10 −2.0000; Cell division protein 48 (CDC48), N-terminal domain CDP-OH_P_transf PF01066.13 0.0000; CDP-alcohol phosphatidyltransferase CS PF04969.8 8.6000; CS domain CSD PF00313.14 −0.3000; ‘Cold-shock’ DNA-binding domain CW_binding_2 PF04122.4 0.0000; Putative cell wall binding repeat 2 Calpain_III PF01067.14 −57.0000; Calpain large subunit, domain III Carbpep_Y_N PF05388.3 2.5000; Carboxypeptidase Y pro-peptide CbiC PF02570.7 25.0000; Precorrin-8X methylmutase Chal_sti_synt_C PF02797.7 −6.1000; Chalcone and stilbene synthases, C-terminal domain Chal_sti_synt_N PF00195.11 −71.3000; Chalcone and stilbene synthases, N-terminal domain CheW PF01584.11 25.0000; CheW-like domain Chorismate_bind PF00425.10 −153.0000; chorismate binding enzyme ClpB_D2-small PF10431.1 21.5000; C-terminal, D2-small domain, of ClpB protein Clp_N PF02861.12 0.0000; Clp amino terminal domain Cna_B PF05738.5 7.0000; Cna protein B-type domain CpcD PF01383.13 3.5000; CpcD/allophycocyanin linker domain Crp PF00325.12 20.7000; Bacterial regulatory proteins, crp family Cupin_2 PF07883.3 19.4000; Cupin domain Cupin_4 PF08007.4 −112.1000; Cupin superfamily protein Cys_Met_Meta_PP PF01053.12 −278.4000; Cys/Met metabolism PLP-dependent enzyme Cytidylate_kin PF02224.10 −51.6000; Cytidylate kinase Cytochrom_C PF00034.13 11.7000; Cytochrome c Cytokin-bind PF09265.2 −143.4000; Cytokinin dehydrogenase 1, FAD and cytokinin binding DAO PF01266.16 −34.9000; FAD dependent oxidoreductase DEAD PF00270.21 7.2000; DEAD/DEAH box helicase DEAD_2 PF06733.7 5.2000; DEAD_2 DHC_N1 PF08385.4 −162.0000; Dynein heavy chain, N-terminal region 1 DHC_N2 PF08393.5 −204.1000; Dynein heavy chain, N-terminal region 2 DHQ_synthase PF01761.12 −119.7000; 3-dehydroquinate synthase DIL PF01843.11 25.0000; DIL domain DNA_pol_lambd_f PF10391.1 25.0000; Fingers domain of DNA polymerase lambda DPPIV_N PF00930.13 −124.2000; Dipeptidyl peptidase IV (DPP IV) N-terminal region DRTGG PF07085.4 −5.3000; DRTGG domain DSPc PF00782.12 −21.8000; Dual specificity phosphatase, catalytic domain DUF1227 PF06777.3 −22.0000; Protein of unknown function (DUF1227) DUF1289 PF06945.5 25.0000; Protein of unknown function (DUF1289) DUF1292 PF06949.3 13.2000; Protein of unknown function (DUF1292) DUF1348 PF07080.3 25.0000; Protein of unknown function (DUF1348) DUF1513 PF07433.3 −172.5000; Protein of unknown function (DUF1513) DUF1644 PF07800.4 −15.6000; Protein of unknown function (DUF1644) DUF1685 PF07939.3 25.0000; Protein of unknown function (DUF1685) DUF1715 PF08215.4 25.0000; Eukaryotic domain of unknown function (DUF1715) DUF1933 PF09147.2 25.0000; Domain of unknown function (DUF1933) DUF1967 PF09269.3 9.0000; Domain of unknown function (DUF1967) DUF1985 PF09331.3 11.9000; Domain of unknown function (DUF1985) DUF442 PF04273.5 −20.4000; Putative phosphatase (DUF442) DUF506 PF04720.4 25.0000; Protein of unknown function (DUF506) DUF537 PF04396.6 −4.6000; Protein of unknown function, DUF537 DUF538 PF04398.4 −24.3000; Protein of unknown function, DUF538 DUF547 PF04784.6 −24.7000; Protein of unknown function, DUF547 DUF568 PF04526.5 25.0000; Protein of unknown function (DUF568) DUF588 PF04535.4 25.0000; Domain of unknown function (DUF588) DUF6 PF00892.12 21.7000; Integral membrane protein DUF6 DUF640 PF04852.4 −18.1000; Protein of unknown function (DUF640) DUF647 PF04884.6 25.0000; Protein of unknown function, DUF647 DUF828 PF05703.3 −53.1000; Plant protein of unknown function (DUF828) DUF836 PF05768.6 2.0000; Glutaredoxin-like domain (DUF836) DZC PF08381.3 15.3000; Disease resistance/zinc finger/chromosome condensation-like region DapB_N PF01113.12 −20.7000; Dihydrodipicolinate reductase, N-terminus DbpA PF03880.7 16.6000; DbpA RNA binding domain DegT_DnrJ_EryC1 PF01041.9 −161.3000; DegT/DnrJ/EryC1/StrS aminotransferase family Dehydrin PF00257.11 −4.4000; Dehydrin Dimerisation PF08100.3 25.0000; Dimerisation domain DnaJ PF00226.23 −8.0000; DnaJ domain Dockerin_1 PF00404.10 24.1000; Dockerin type I repeat DoxX PF07681.4 0.3000; DoxX DpmII PF04556.4 25.0000; DpmII restriction endonuclease Dynein_heavy PF03028.7 −455.0000; Dynein heavy chain EAL PF00563.12 −16.4000; EAL domain EFG_C PF00679.16 6.0000; Elongation factor G C-terminus EFG_IV PF03764.10 25.0000; Elongation factor G, domain IV EIN3 PF04873.5 −192.4000; Ethylene insensitive 3 ENT PF03735.6 14.7000; ENT domain Exo_endo_phos PF03372.15 11.0000; Endonuclease/Exonuclease/phosphatase family Exostosin PF03016.7 −53.2000; Exostosin family ExsB PF06508.5 −91.9000; ExsB F-box PF00646.25 14.7000; F-box domain F420_oxidored PF03807.9 −34.5000; NADP oxidoreductase coenzyme F420- dependent FAD-oxidase_C PF02913.11 −36.0000; FAD linked oxidases, C-terminal domain FAD_binding_1 PF00667.12 −79.0000; FAD binding domain FAD_binding_3 PF01494.11 −136.6000; FAD binding domain FAD_binding_4 PF01565.15 −8.1000; FAD binding domain FAD_binding_5 PF00941.13 −57.4000; FAD binding domain in molybdopterin dehydrogenase FAD_binding_6 PF00970.16 −4.9000; Oxidoreductase FAD-binding domain FAD_binding_8 PF08022.4 −10.4000; FAD-binding domain FAD_binding_9 PF08021.3 −22.9000; Siderophore-interacting FAD-binding domain FA_desaturase PF00487.16 0.2000; Fatty acid desaturase FCD PF07729.4 29.0000; FCD domain FHA PF00498.18 25.0000; FHA domain FLYWCH PF04500.8 12.8000; FLYWCH zinc finger domain FMN_dh PF01070.10 −203.2000; FMN-dependent dehydrogenase FTCD PF02971.6 25.0000; Formiminotransferase domain FTCD_C PF04961.4 −69.6000; Formiminotransferase-cyclodeaminase FTCD_N PF07837.4 −110.1000; Formiminotransferase domain, N-terminal subdomain FYRC PF05965.6 17.1000; F/Y rich C-terminus FYRN PF05964.6 16.3000; F/Y-rich N-terminus FYVE PF01363.13 3.9000; FYVE zinc finger F_bP_aldolase PF01116.12 −157.6000; Fructose-bisphosphate aldolase class-II Fe-ADH PF00465.11 −189.8000; Iron-containing alcohol dehydrogenase Fe—S_biosyn PF01521.12 −0.4000; Iron-sulphur cluster biosynthesis FeoB_C PF07664.4 16.6000; Ferrous iron transport protein B C terminus FeoB_N PF02421.10 −0.4000; Ferrous iron transport protein B Fer2 PF00111.19 11.2000; 2Fe—2S iron-sulfur cluster binding domain Fer2_BFD PF04324.7 16.0000; BFD-like [2Fe—2S] binding domain Fer4 PF00037.19 15.5000; 4Fe—4S binding domain Ferric_reduct PF01794.11 −7.0000; Ferric reductase like transmembrane component Flavodoxin_1 PF00258.17 6.3000; Flavodoxin FleQ PF06490.3 −14.4000; Flagellar regulatory protein FleQ Form_Nir_trans PF01226.9 −131.5000; Formate/nitrite transporter FrhB_FdhB_C PF04432.5 −42.4000; Coenzyme F420 hydrogenase/dehydrogenase, beta subunit C terminus FrhB_FdhB_N PF04422.5 4.6000; Coenzyme F420 hydrogenase/dehydrogenase, beta subunit N terminus FtsH_ext PF06480.7 6.8000; FtsH Extracellular Fumble PF03630.6 25.0000; Fumble GAF PF01590.18 23.0000; GAF domain GATA PF00320.19 28.5000; GATA zinc finger GATase_2 PF00310.13 −91.6000; Glutamine amidotransferases class-II GCC2_GCC3 PF07699.5 30.0000; GCC2 and GCC3 GDPD PF03009.9 −18.0000; Glycerophosphoryl diester phosphodiesterase family GFO_IDH_MocA PF01408.14 1.0000; Oxidoreductase family, NAD-binding Rossmann fold GGDEF PF00990.13 −45.0000; GGDEF domain GH3 PF03321.5 −336.0000; GH3 auxin-responsive promoter GRAS PF03514.6 −181.6000; GRAS family transcription factor GSPII_E PF00437.12 −98.1000; Type II/IV secretion system protein GSPII_E_N PF05157.7 5.7000; GSPII_E N-terminal domain GTP1_OBG PF01018.14 −56.2000; GTP1/OBG GTP_EFTU PF00009.19 8.0000; Elongation factor Tu GTP binding domain GTP_EFTU_D2 PF03144.17 25.0000; Elongation factor Tu domain 2 GTP_EFTU_D3 PF03143.9 14.3000; Elongation factor Tu C-terminal domain GUCT PF08152.4 25.0000; GUCT (NUC152) domain GXGXG PF01493.11 5.0000; GXGXG motif Gal_Lectin PF02140.10 25.0000; Galactose binding lectin domain GatB_Yqey PF02637.10 −28.5000; GatB domain Gate PF07670.6 29.5000; Nucleoside recognition GcpE PF04551.6 −312.3000; GcpE protein GerE PF00196.11 6.1000; Bacterial regulatory proteins, luxR family Gln-synt_C PF00120.16 −124.0000; Glutamine synthetase, catalytic domain Gln-synt_N PF03951.11 9.0000; Glutamine synthetase, beta-Grasp domain Globin PF00042.14 −7.3000; Globin Glt_symporter PF03616.6 −272.0000; Sodium/glutamate symporter Glu_syn_central PF04898.6 25.0000; Glutamate synthase central domain Glu_synthase PF01645.9 −198.4000; Conserved region in glutamate synthase Glutaredoxin PF00462.16 17.2000; Glutaredoxin Glyco_hydro_1 PF00232.10 −301.8000; Glycosyl hydrolase family 1 Glyco_hydro_28 PF00295.9 −97.0000; Glycosyl hydrolases family 28 Glyco_hydro_35 PF01301.11 −184.5000; Glycosyl hydrolases family 35 Glyco_hydro_42 PF02449.7 −230.8000; Beta-galactosidase Glyco_transf_20 PF00982.13 −243.6000; Glycosyltransferase family 20 Glyco_transf_9 PF01075.9 −62.2000; Glycosyltransferase family 9 (heptosyltransferase) Glycos_transf_1 PF00534.12 −7.3000; Glycosyl transferases group 1 Glycos_transf_2 PF00535.18 17.6000; Glycosyl transferase family 2 Got1 PF04178.4 25.0000; Got1-like family Gp_dh_C PF02800.12 −64.1000; Glyceraldehyde 3-phosphate dehydrogenase, C- terminal domain Guanylate_cyc PF00211.12 −23.7000; Adenylate and Guanylate cyclase catalytic domain HALZ PF02183.10 18.1000; Homeobox associated leucine zipper HAMP PF00672.17 19.8000; HAMP domain HATPase_c PF02518.18 22.4000; Histidine kinase-, DNA gyrase B-, and HSP90- like ATPase HD PF01966.14 17.8000; HD domain HD-ZIP_N PF04618.4 25.0000; HD-ZIP protein N terminus HDAC_interact PF08295.4 −17.1000; Histone deacetylase (HDAC) interacting HEPN PF05168.6 0.1000; HEPN domain HIG_1_N PF04588.5 −5.2000; Hypoxia induced protein conserved region HLH PF00010.18 8.3000; Helix-loop-helix DNA-binding domain HMG_box PF00505.11 4.1000; HMG (high mobility group) box HPPK PF01288.12 25.0000; 7,8-dihydro-6-hydroxymethylpterin- pyrophosphokinase (HPPK) HR1 PF02185.8 0.0000; Hr1 repeat HR_lesion PF05514.3 25.0000; HR-like lesion-inducing HSP20 PF00011.13 13.0000; Hsp20/alpha crystallin family HSP70 PF00012.12 −286.0000; Hsp70 protein HTH_3 PF01381.14 27.7000; Helix-turn-helix HTH_6 PF01418.9 0.0000; Helix-turn-helix domain, rpiR family HTH_8 PF02954.11 17.3000; Bacterial regulatory protein, Fis family HTH_AraC PF00165.15 22.0000; Bacterial regulatory helix-turn-helix proteins, AraC family Helicase_C PF00271.23 6.9000; Helicase conserved C-terminal domain HemolysinCabind PF00353.11 5.0000; Hemolysin-type calcium-binding repeat (2 copies) HisKA PF00512.17 22.1000; His Kinase A (phosphoacceptor) domain HisKA_3 PF07730.5 30.0000; Histidine kinase His_kinase PF06580.5 25.0000; Histidine kinase Homeobox PF00046.21 −4.1000; Homeobox domain Homoserine_dh PF00742.11 −5.0000; Homoserine dehydrogenase Hpt PF01627.15 25.0000; Hpt domain Hydrolase PF00702.18 13.6000; haloacid dehalogenase-like hydrolase Hydrolase_3 PF08282.4 −64.8000; haloacid dehalogenase-like hydrolase IGPD PF00475.10 25.0000; Imidazoleglycerol-phosphate dehydratase IQ PF00612.19 18.3000; IQ calmodulin-binding motif IlvN PF07991.4 −75.8000; Acetohydroxy acid isomeroreductase, catalytic domain K-box PF01486.9 0.0000; K-box region KA1 PF02149.11 25.0000; Kinase associated domain 1 KH_2 PF07650.9 5.0000; KH domain KOW PF00467.21 29.1000; KOW motif KR PF08659.2 −74.3000; KR domain Kelch_1 PF01344.17 11.7000; Kelch motif Kelch_2 PF07646.7 14.0000; Kelch motif Kinesin PF00225.15 −135.0000; Kinesin motor domain LIM PF00412.14 0.0000; LIM domain LRRNT_2 PF08263.4 18.6000; Leucine rich repeat N-terminal domain LRR_1 PF00560.25 10.9000; Leucine Rich Repeat LRR_2 PF07723.5 14.5000; Leucine Rich Repeat LRR_3 PF07725.4 20.0000; Leucine Rich Repeat Lact-deh-memb PF09330.3 25.0000; D-lactate dehydrogenase, membrane binding Lactamase_B PF00753.19 24.6000; Metallo-beta-lactamase superfamily Lectin_legB PF00139.11 −110.1000; Legume lectin domain Lipase_GDSL PF00657.14 13.9000; GDSL-like Lipase/Acylhydrolase Lipoprotein_9 PF03180.6 −121.0000; NLPA lipoprotein LisH PF08513.3 20.7000; LisH LysM PF01476.12 20.0000; LysM domain MBD PF01429.11 12.9000; Methyl-CpG binding domain MCD PF05292.3 25.0000; Malonyl-CoA decarboxylase (MCD) MCPsignal PF00015.13 −88.3000; Methyl-accepting chemotaxis protein (MCP) signaling domain MEKHLA PF08670.3 −59.7000; MEKHLA domain MFS_1 PF07690.8 23.5000; Major Facilitator Superfamily MIT PF04212.10 18.3000; MIT (microtubule interacting and transport) domain MMR_HSR1 PF01926.15 31.2000; GTPase of unknown function MOFRL PF05161.5 25.0000; MOFRL family MORN PF02493.12 0.0000; MORN repeat MOSC PF03473.9 25.0000; MOSC domain MOSC_N PF03476.8 25.0000; MOSC N-terminal beta barrel domain Malic_M PF03949.7 −143.9000; Malic enzyme, NAD binding domain MaoC_dehydratas PF01575.11 −10.9000; MaoC like domain Metal_hydrol PF10118.1 −74.9000; Predicted metal-dependent hydrolase Metallophos PF00149.20 22.0000; Calcineurin-like phosphoesterase MethyltransfD12 PF02086.7 −49.4000; D12 class N6 adenine-specific DNA methyltransferase Methyltransf_11 PF08241.4 24.4000; Methyltransferase domain Methyltransf_12 PF08242.4 23.0000; Methyltransferase domain Methyltransf_16 PF10294.1 −43.0000; Putative methyltransferase Methyltransf_2 PF00891.10 −103.8000; O-methyltransferase Miro PF08477.5 10.8000; Miro-like protein Mito_carr PF00153.19 0.0000; Mitochondrial carrier protein Molybdop_Fe4S4 PF04879.8 13.6000; Molybdopterin oxidoreductase Fe4S4 domain Molybdopterin PF00384.14 −50.0000; Molybdopterin oxidoreductase Molydop_binding PF01568.13 1.1000; Molydopterin dinucleotide binding domain Monooxygenase_B PF04744.4 25.0000; Monooxygenase subunit B protein Mur_ligase PF01225.17 4.9000; Mur ligase family, catalytic domain Mur_ligase_C PF02875.13 8.0000; Mur ligase family, glutamate ligase domain Mur_ligase_M PF08245.4 −44.7000; Mur ligase middle domain Myb_DNA-binding PF00249.23 21.7000; Myb-like DNA-binding domain Myosin_N PF02736.11 9.0000; Myosin N-terminal SH3-like domain Myosin_head PF00063.13 −372.4000; Myosin head (motor domain) N6_N4_Mtase PF01555.10 −36.4000; DNA methylase NAD_Gly3P_dh_N PF01210.15 −44.0000; NAD-dependent glycerol-3-phosphate dehydrogenase N-terminus NAD_binding_1 PF00175.13 −3.9000; Oxidoreductase NAD-binding domain NAD_binding_2 PF03446.7 −63.5000; NAD binding domain of 6-phosphogluconate dehydrogenase NAD_binding_3 PF03447.8 −1.7000; Homoserine dehydrogenase, NAD binding domain NAD_binding_6 PF08030.4 −23.6000; Ferric reductase NAD binding domain NAF PF03822.6 4.5000; NAF domain NAGLU PF05089.4 25.0000; Alpha-N-acetylglucosaminidase (NAGLU) NAM PF02365.7 −19.0000; No apical meristem (NAM) protein NB-ARC PF00931.14 −79.5000; NB-ARC domain NIR_SIR PF01077.14 −18.7000; Nitrite and sulphite reductase 4Fe—4S domain NIR_SIR_ferr PF03460.9 2.4000; Nitrite/Sulfite reductase ferredoxin-like half domain NLE PF08154.4 14.0000; NLE (NUC135) domain NMT1 PF09084.3 20.0000; NMT1/THI5 like NOG1 PF06858.6 50.0000; Nucleolar GTP-binding protein 1 (NOG1) NTP_transf_2 PF01909.15 18.8000; Nucleotidyltransferase domain NTP_transferase PF00483.15 −39.9000; Nucleotidyl transferase NUDIX PF00293.20 0.2000; NUDIX domain Ndr PF03096.6 −82.0000; Ndr family NifU PF01106.9 −5.6000; NifU-like domain Nitroreductase PF00881.16 14.2000; Nitroreductase family Nramp PF01566.10 −171.4000; Natural resistance-associated macrophage protein OB_RNB PF08206.3 4.1000; Ribonuclease B OB domain OKR_DC_1 PF01276.12 −206.6000; Orn/Lys/Arg decarboxylase, major domain OKR_DC_1_C PF03711.7 −52.0000; Orn/Lys/Arg decarboxylase, C-terminal domain OKR_DC_1_N PF03709.7 7.0000; Orn/Lys/Arg decarboxylase, N-terminal domain OMPdecase PF00215.16 −45.6000; Orotidine 5′-phosphate decarboxylase/HUMPS family OPT PF03169.7 −233.0000; OPT oligopeptide transporter protein OpuAC PF04069.4 −45.4000; Substrate binding domain of ABC-type glycine betaine transport system Orn_Arg_deC_N PF02784.8 −76.0000; Pyridoxal-dependent decarboxylase, pyridoxal binding domain Orn_DAP_Arg_deC PF00278.14 6.7000; Pyridoxal-dependent decarboxylase, C-terminal sheet domain P-II PF00543.14 −29.0000; Nitrogen regulatory protein P-II PAH PF02671.13 25.0000; Paired amphipathic helix repeat PAPS_reduct PF01507.11 −53.4000; Phosphoadenosine phosphosulfate reductase family PAS PF00989.16 0.0000; PAS fold PAS_2 PF08446.3 −2.1000; PAS fold PAS_3 PF08447.3 17.2000; PAS fold PAS_4 PF08448.2 16.4000; PAS fold PB1 PF00564.16 12.3000; PB1 domain PBP PF01161.12 −20.6000; Phosphatidylethanolamine-binding protein PC_rep PF01851.14 0.0000; Proteasome/cyclosome repeat PGI PF00342.11 −146.0000; Phosphoglucose isomerase PGM_PMM_I PF02878.8 −37.5000; Phosphoglucomutase/phosphomannomutase, alpha/beta/alpha domain I PGM_PMM_II PF02879.8 −20.0000; Phosphoglucomutase/phosphomannomutase, alpha/beta/alpha domain II PHD PF00628.21 26.2000; PHD-finger PH_2 PF08458.2 −6.6000; Plant pleckstrin homology-like region PLAC8 PF04749.9 −1.1000; PLAC8 family PMEI PF04043.7 25.0000; Plant invertase/pectin methylesterase inhibitor PMSR PF01625.13 −32.7000; Peptide methionine sulfoxide reductase PNK3P PF08645.3 −67.6000; Polynucleotide kinase 3 phosphatase PNTB PF02233.8 −341.0000; NAD(P) transhydrogenase beta subunit POX PF07526.3 −39.4000; Associated with HOX PP-binding PF00550.17 25.3000; Phosphopantetheine attachment site PP2C PF00481.13 −36.3000; Protein phosphatase 2C PPR PF01535.12 1.6000; PPR repeat PRP4 PF08799.3 25.0000; pre-mRNA processing factor 4 (PRP4) like PSCyt1 PF07635.3 50.0000; Planctomycete cytochrome C PTA_PTB PF01515.11 −165.5000; Phosphate acetyl/butaryl transferase PTR2 PF00854.13 −50.0000; POT family PWWP PF00855.9 0.3000; PWWP domain Pantoate_ligase PF02569.7 −106.0000; Pantoate-beta-alanine ligase Pantoate_transf PF02548.7 −93.0000; Ketopantoate hydroxymethyltransferase PduL PF06130.4 25.0000; Propanediol utilisation protein PduL Pectinesterase PF01095.11 −222.9000; Pectinesterase PepSY_TM PF03929.8 11.9000; PepSY-associated TM helix Peptidase_C14 PF00656.14 −22.5000; Caspase domain Peptidase_C2 PF00648.13 −183.5000; Calpain family cysteine protease Peptidase_M3 PF01432.12 −146.0000; Peptidase family M3 Peptidase_M41 PF01434.10 −139.8000; Peptidase family M41 Peptidase_S10 PF00450.14 −198.0000; Serine carboxypeptidase Peptidase_S24 PF00717.15 10.0000; Peptidase S24-like Pericardin_rpt PF07054.3 0.0000; Pericardin like repeat Peripla_BP_1 PF00532.13 −44.9000; Periplasmic binding proteins and sugar binding domain of the LacI family Phage_fiber PF03335.5 0.0000; Phage tail fibre repeat Phytochrome PF00360.12 13.0000; Phytochrome region PilZ PF07238.6 19.6000; PilZ domain Pkinase PF00069.17 70.3000; Protein kinase domain Pkinase_C PF00433.16 15.3000; Protein kinase C terminal domain Pkinase_Tyr PF07714.9 −155.0000; Protein tyrosine kinase Plant_zn_clust PF10533.1 25.0000; Plant zinc cluster domain Plus-3 PF03126.10 25.0000; Plus-3 domain Pollen_allerg_1 PF01357.13 17.2000; Pollen allergen Pribosyltran PF00156.19 20.0000; Phosphoribosyl transferase domain Proteasome PF00227.18 −36.7000; Proteasome A-type and B-type PseudoU_synth_2 PF00849.14 −18.0000; RNA pseudouridylate synthase Pterin_bind PF00809.14 −53.7000; Pterin binding enzyme Pyr_redox PF00070.19 5.0000; Pyridine nucleotide-disulphide oxidoreductase Pyr_redox_2 PF07992.6 −20.0000; Pyridine nucleotide-disulphide oxidoreductase Pyridox_oxidase PF01243.12 17.1000; Pyridoxamine 5′-phosphate oxidase Pyrophosphatase PF00719.11 −41.0000; Inorganic pyrophosphatase RCC1 PF00415.10 21.2000; Regulator of chromosome condensation (RCC1) RIO1 PF01163.14 −89.1000; RIO1 family RNA_Me_trans PF04252.5 25.0000; Predicted SAM-dependent RNA methyltransferase RNA_pol_L PF01193.16 16.9000; RNA polymerase Rpb3/Rpb11 dimerisation domain RRM_1 PF00076.14 17.9000; RNA recognition motif. (a.k.a. RRM, RBD, or RNP domain) RVT_1 PF00078.19 32.0000; Reverse transcriptase (RNA-dependent DNA polymerase) RVT_2 PF07727.6 −180.0000; Reverse transcriptase (RNA-dependent DNA polymerase) RWP-RK PF02042.7 25.0000; RWP-RK domain RecA PF00154.13 −100.6000; recA bacterial DNA recombination protein Reg_prop PF07494.3 5.4000; Two component regulator propeller ResIII PF04851.7 0.8000; Type III restriction enzyme, res subunit Response_reg PF00072.16 4.1000; Response regulator receiver domain Rho_N PF07498.4 31.0000; Rho termination factor, N-terminal domain Rhodanese PF00581.12 25.0000; Rhodanese-like domain Rhomboid PF01694.14 −1.2000; Rhomboid family Ribosomal_L21p PF00829.13 −23.0000; Ribosomal prokaryotic L21 protein Ribosomal_S19 PF00203.13 11.0000; Ribosomal protein S19 Ribosomal_S9 PF00380.11 −24.0000; Ribosomal protein S9/S16 Rieske PF00355.18 −7.0000; Rieske [2Fe—2S] domain RolB_RolC PF02027.9 25.0000; RolB/RolC glucosidase family Rrf2 PF02082.12 −17.5000; Transcriptional regulator RrnaAD PF00398.12 −73.3000; Ribosomal RNA adenine dimethylase Rubredoxin PF00301.12 14.0000; Rubredoxin S-AdoMet_synt_M PF02772.8 25.0000; S-adenosylmethionine synthetase, central domain S4 PF01479.17 15.0000; S4 domain S6PP PF05116.5 −113.7000; Sucrose-6F-phosphate phosphohydrolase SAM_1 PF00536.22 11.3000; SAM domain (Sterile alpha motif) SAM_2 PF07647.9 0.0000; SAM domain (Sterile alpha motif) SBP PF03110.6 25.0000; SBP domain SBP_bac_3 PF00497.12 16.6000; Bacterial extracellular solute-binding proteins, family 3 SCP PF00188.18 10.0000; SCP-like extracellular protein SET PF00856.20 23.5000; SET domain SH3BGR PF04908.7 −12.0000; SH3-binding, glutamic acid-rich protein SIS PF01380.14 0.0000; SIS domain SLA_LP_auto_ag PF05889.5 −170.7000; Soluble liver antigen/liver pancreas antigen (SLA/LP autoantigen) SMC_N PF02463.11 −95.8000; RecF/RecN/SMC N terminal domain SOUL PF04832.4 25.0000; SOUL heme-binding protein SPRY PF00622.20 10.0000; SPRY domain SRF-TF PF00319.10 11.0000; SRF-type transcription factor (DNA-binding and dimerisation domain) SSF PF00474.9 −162.3000; Sodium:solute symporter family START PF01852.11 −20.7000; START domain STAS PF01740.13 0.0000; STAS domain SbmA_BacA PF05992.4 −145.6000; SbmA/BacA-like family SelR PF01641.10 −66.5000; SelR domain Shikimate_DH PF01488.12 −4.4000; Shikimate/quinate 5-dehydrogenase Shugoshin_C PF07557.3 25.0000; Shugoshin C terminus Sigma54_activat PF00158.18 −95.0000; Sigma-54 interaction domain Sina PF03145.8 −48.4000; Seven in absentia protein family SirA PF01206.9 7.0000; SirA-like protein SpoIIE PF07228.4 −13.7000; Stage II sporulation protein E (SpoIIE) Spore_permease PF03845.5 −134.0000; Spore germination protein Ssl1 PF04056.6 −151.8000; Ssl1-like Suc_Fer-like PF06999.4 −42.4000; Sucrase/ferredoxin-like Sulfate_transp PF00916.12 −131.5000; Sulfate transporter family Synaptobrevin PF00957.13 25.0000; Synaptobrevin TAT_signal PF10518.1 22.9000; TAT (twin-arginine translocation) pathway signal sequence TFIID-31 kDa PF02291.7 25.0000; Transcription initiation factor IID, 31 kD subunit TFIID_90 kDa PF04494.7 25.0000; WD40 associated region in TFIID subunit TGS PF02824.13 5.6000; TGS domain TIR PF01582.12 43.5000; TIR domain TPP_enzyme_C PF02775.13 19.7000; Thiamine pyrophosphate enzyme, C-terminal TPP binding domain TPP_enzyme_M PF00205.14 −8.1000; Thiamine pyrophosphate enzyme, central domain TPP_enzyme_N PF02776.10 −70.0000; Thiamine pyrophosphate enzyme, N-terminal TPP binding domain TPR_1 PF00515.20 16.1000; Tetratricopeptide repeat TPR_2 PF07719.9 23.9000; Tetratricopeptide repeat Thg1 PF04446.4 25.0000; tRNAHis guanylyltransferase ThiS PF02597.12 19.3000; ThiS family Thioesterase PF00975.12 −10.5000; Thioesterase domain Thiolase_C PF02803.10 −30.7000; Thiolase, C-terminal domain Thiolase_N PF00108.15 −129.5000; Thiolase, N-terminal domain Thioredoxin PF00085.12 −16.6000; Thioredoxin Thymidylate_kin PF02223.9 2.3000; Thymidylate kinase Torsin PF06309.3 −158.6000; Torsin TraB PF01963.9 −62.0000; TraB family Trans_reg_C PF00486.20 25.0000; Transcriptional regulatory protein, C terminal Transaldolase PF00923.11 −49.0000; Transaldolase Trehalose_PPase PF02358.8 −49.4000; Trehalose-phosphatase TrkA_C PF02080.13 25.0000; TrkA-C domain TrkA_N PF02254.10 4.7000; TrkA-N domain Trp_Tyr_perm PF03222.5 −232.6000; Tryptophan/tyrosine permease family Trp_halogenase PF04820.6 −283.2000; Tryptophan halogenase Tub PF01167.10 −98.0000; Tub family Tubulin PF00091.17 −48.7000; Tubulin/FtsZ family, GTPase domain Tup_N PF08581.2 25.0000; Tup N-terminal U-box PF04564.7 −7.6000; U-box domain U3_snoRNA_C PF09384.2 25.0000; U3 small nucleolar RNA C terminal UBA PF00627.23 22.4000; UBA/TS-N domain UBA_2 PF08587.3 25.0000; Ubiquitin associated domain (UBA) UDPG_MGDP_dh_N PF03721.6 −67.4000; UDP-glucose/GDP-mannose dehydrogenase family, NAD binding domain UIM PF02809.12 16.6000; Ubiquitin interaction motif UVR PF02151.11 16.0000; UvrB/uvrC motif Usp PF00582.18 22.3000; Universal stress protein family Utp21 PF04192.4 25.0000; Utp21 specific WD40 associated putative domain Vps4_C PF09336.2 −4.6000; Vps4 C terminal oligomerisation domain WD40 PF00400.24 21.5000; WD domain, G-beta repeat WRKY PF03106.7 −6.7000; WRKY DNA-binding domain WW PF00397.18 0.0000; WW domain XH PF03469.6 25.0000; XH domain XS PF03468.6 25.0000; XS domain XYPPX PF02162.9 0.7000; XYPPX repeat Xan_ur_permease PF00860.12 −146.7000; Permease family YLP PF02757.9 2.7000; YLP motif Y_Y_Y PF07495.5 5.0000; Y_Y_Y domain Zn_clus PF00172.10 12.0000; Fungal Zn(2)-Cys(6) binuclear cluster domain adh_short PF00106.17 −40.2000; short chain dehydrogenase bZIP_1 PF00170.13 24.8000; bZIP transcription factor bZIP_2 PF07716.7 15.0000; Basic region leucine zipper cNMP_binding PF00027.21 20.6000; Cyclic nucleotide-binding domain eIF-5a PF01287.12 9.6000; Eukaryotic initiation factor 5A hypusine, DNA- binding OB fold eIF2A PF08662.3 −71.9000; Eukaryotic translation initiation factor eIF2A eRF1_1 PF03463.7 −12.9000; eRF1 domain 1 eRF1_2 PF03464.7 3.6000; eRF1 domain 2 eRF1_3 PF03465.7 1.4000; eRF1 domain 3 Efhand PF00036.24 21.7000; EF hand fn3 PF00041.13 7.2000; Fibronectin type III domain mTERF PF02536.6 −60.0000; mTERF Malic PF00390.11 25.0000; Malic enzyme, N-terminal domain oligo_HPY PF08352.4 2.6000; Oligopeptide/dipeptide transporter, C-terminal region p450 PF00067.14 −105.0000; Cytochrome P450 Peroxidase PF00141.15 −10.0000; Peroxidase Rve PF00665.18 32.0000; Integrase core domain tRNA-synt_1c PF00749.13 −130.2000; tRNA synthetases class I (E and Q), catalytic domain tRNA-synt_1c_C PF03950.10 25.0000; tRNA synthetases class I (E and Q), anti-codon binding domain tRNA-synt_1g PF09334.3 −272.5000; tRNA synthetases class I (M) tRNA_synt_1c_R1 PF04558.7 25.0000; Glutaminyl-tRNA synthetase, non-specific RNA binding region part 1 tRNA_synt_1c_R2 PF04557.7 25.0000; Glutaminyl-tRNA synthetase, non-specific RNA binding region part 2 Ubiquitin PF00240.15 19.4000; Ubiquitin family zf-B_box PF00643.16 15.3000; B-box zinc finger zf-C3HC4 PF00097.17 16.0000; Zinc finger, C3HC4 type (RING finger) zf-CCCH PF00642.16 0.0000; Zinc finger C-x8-C-x5-C-x3-H type (and similar) zf-CCHC PF00098.15 17.9000; Zinc knuckle zf-CDGSH PF09360.2 10.4000; Iron-binding zinc finger CDGSH type zf-CW PF07496.7 25.0000; CW-type Zinc Finger zf-Dof PF02701.7 25.0000; Dof domain, zinc finger zf-GRF PF06839.4 22.7000; GRF zinc finger zf-RanBP PF00641.10 23.0000; Zn-finger in Ran binding protein and others zf-TRAF PF02176.10 18.6000; TRAF-type zinc finger zf-dskA_traR PF01258.9 21.0000; Prokaryotic dksA/traR C4-type zinc finger 

1. A recombinant DNA construct comprising a polynucleotide encoding a protein having a CCT Pfam domain scoring above the Pfam gathering cutoff value of 25.000.
 2. The recombinant DNA construct of claim 1 wherein said protein has an amino acid sequence having at least 90% identity over at least 90% of the length of a reference sequence having the amino acid sequence of SEQ ID NO: 374 when said amino acid sequence is aligned with said reference sequence.
 3. A recombinant DNA construct comprising a promoter that is functional in a plant cell and that is operably linked to a polynucleotide that, when expressed in a plant cell: (a) encodes a protein: i) having an amino acid sequence selected from the group consisting of SEQ ID NO: 308, 310, 312-315, 317-323, 325-343, 345, 347-349, 352-354, 356, 358-359, 366-372, 374-383, 389-392, 394, 396, 401-403, 405-412, 414, 417-424, 427-453, 455-473, 475, 488-501, 503-517, 519-531, 533-540, 542-543, and 546-614; ii) having an amino acid sequence having at least 90% identity over at least 90% of a reference sequence selected from the group consisting of SEQ ID NO: 308, 310, 312-315, 317-323, 325-343, 345, 347-349, 352-354, 356, 358-359, 366-372, 374-383, 389-392, 394, 396, 401-403, 405-412, 414, 417-424, 427-453, 455-473, 475, 488-501, 503-517, 519-531, 533-540, 542-543, and 546-614 when said amino acid sequence is aligned to said reference sequence; or iii) that is a homolog of a protein with an amino acid sequence selected from the group consisting of SEQ ID NO: 308, 310, 312-315, 317-323, 325-343, 345, 347-349, 352-354, 356, 358-359, 366-372, 374-383, 389-392, 394, 396, 401-403, 405-412, 414, 417-424, 427-453, 455-473, 475, 488-501, 503-517, 519-531, 533-540, 542-543, and 546-614; or (b) is transcribed into an RNA molecule that suppresses the level of an endogenous protein in said plant cell wherein said endogenous protein has an amino acid sequence selected from the group consisting of SEQ ID NO: 309, 311, 316, 324, 344, 346, 350-351, 355, 357, 360-365, 373, 384-388, 393, 395, 397-400, 404, 413, 415-416, 425-426, 454, 474, 476-487, 502, 518, 532, 541, or 544-545 or is a homolog thereof; wherein said construct is stably integrated into plant chromosomal DNA.
 4. A transgenic plant cell comprising the recombinant DNA construct of claim 3 wherein said plant cell is in a plant selected by screening a population of transgenic plants that have been transformed with said construct for an enhanced trait as compared to control plants; and wherein said enhanced trait is enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein or enhanced seed oil.
 5. The plant cell of claim 4 further comprising DNA expressing a protein that provides tolerance from exposure to an herbicide comprising an agent applied at levels that are lethal to a wild type of said plant cell.
 6. The plant cell of claim 5 wherein the agent of said herbicide is a glyphosate, dicamba, or glufosinate compound.
 7. A transgenic plant comprising a plurality of plant cells of claim
 4. 8. The transgenic plant of claim 7 which is homozygous for said recombinant DNA.
 9. A transgenic seed comprising a plurality of plant cells of claim
 4. 10. The transgenic seed of claim 9 from a corn, soybean, cotton, canola, alfalfa, wheat, rice, sugarcane, or sugar beet plant.
 11. Grain comprising transgenic seed identifiable by the recombinant DNA construct of claim
 3. 12. Seed meal produced from transgenic seed identifiable by the recombinant DNA construct of claim
 3. 13. A transgenic pollen grain comprising a haploid derivative of a plant cell nucleus having a chromosome comprising the recombinant DNA construct of claim
 3. 14. A method for manufacturing non-natural, transgenic seed that can be used to produce a crop of transgenic plants with an enhanced trait resulting from expression of the stably-integrated, recombinant DNA construct of claim 3, said method comprising: (a) screening a population of plants for said enhanced trait and said recombinant DNA, wherein individual plants in said population exhibit said trait at a level less than, essentially the same as or greater than the level that said trait is exhibited in control plants which do not contain said recombinant DNA, wherein said enhanced trait is selected from the group of enhanced traits consisting of enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil; (b) selecting from said population one or more plants that exhibit said trait at a level greater than the level that said trait is exhibited in control plants, and (c) collecting seed from selected plant from step b.
 15. The method of claim 14 wherein said method for manufacturing said transgenic seed further comprises: (a) verifying that said recombinant DNA is stably integrated in said selected plants, and (b) analyzing tissue of said selected plant to determine the expression or suppression of a protein having the function of a protein having an amino acid sequence selected from the group consisting of one of SEQ ID NOs:308-614.
 16. The method of claim 15 wherein said seed is corn, soybean, cotton, canola, alfalfa, wheat, rice, sugarcane, or sugar beet seed.
 17. A method of producing hybrid corn seed comprising: (a) acquiring hybrid corn seed from an herbicide tolerant corn plant which also has the stably-integrated, recombinant DNA construct of claim 3; (b) producing corn plants from said hybrid corn seed, wherein a fraction of the plants produced from said hybrid corn seed is homozygous for said recombinant DNA, a fraction of the plants produced from said hybrid corn seed is hemizygous for said recombinant DNA, and a fraction of the plants produced from said hybrid corn seed has none of said recombinant DNA; (c) selecting corn plants which are homozygous and hemizygous for said recombinant DNA by treating with an herbicide; (d) collecting seed from herbicide-treated-surviving corn plants and planting said seed to produce further progeny corn plants; (e) repeating steps (c) and (d) at least once to produce an inbred corn line; and (f) crossing said inbred corn line with a second corn line to produce hybrid seed. 